78 results on '"Minn AJ"'
Search Results
2. Combination anti-PD-1 and anti-CTLA-4 therapy generates waves of clonal responses that include progenitor-exhausted CD8 + T cells.
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Wang K, Coutifaris P, Brocks D, Wang G, Azar T, Solis S, Nandi A, Anderson S, Han N, Manne S, Kiner E, Sachar C, Lucas M, George S, Yan PK, Kier MW, Laughlin AI, Kothari S, Giles J, Mathew D, Ghinnagow R, Alanio C, Flowers A, Xu W, Tenney DJ, Xu X, Amaravadi RK, Karakousis GC, Schuchter LM, Buggert M, Oldridge D, Minn AJ, Blank C, Weber JS, Mitchell TC, Farwell MD, Herati RS, and Huang AC
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- Humans, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Antineoplastic Combined Chemotherapy Protocols pharmacology, Receptors, Antigen, T-Cell immunology, Receptors, Antigen, T-Cell metabolism, Female, Single-Cell Analysis methods, Male, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes drug effects, Programmed Cell Death 1 Receptor antagonists & inhibitors, Programmed Cell Death 1 Receptor immunology, Melanoma drug therapy, Melanoma immunology, Immune Checkpoint Inhibitors pharmacology, Immune Checkpoint Inhibitors therapeutic use, CTLA-4 Antigen antagonists & inhibitors, CTLA-4 Antigen immunology
- Abstract
Combination checkpoint blockade with anti-PD-1 and anti-CTLA-4 antibodies has shown promising efficacy in melanoma. However, the underlying mechanism in humans remains unclear. Here, we perform paired single-cell RNA and T cell receptor (TCR) sequencing across time in 36 patients with stage IV melanoma treated with anti-PD-1, anti-CTLA-4, or combination therapy. We develop the algorithm Cyclone to track temporal clonal dynamics and underlying cell states. Checkpoint blockade induces waves of clonal T cell responses that peak at distinct time points. Combination therapy results in greater magnitude of clonal responses at 6 and 9 weeks compared to single-agent therapies, including melanoma-specific CD8
+ T cells and exhausted CD8+ T cell (TEX ) clones. Focused analyses of TEX identify that anti-CTLA-4 induces robust expansion and proliferation of progenitor TEX , which synergizes with anti-PD-1 to reinvigorate TEX during combination therapy. These next generation immune profiling approaches can guide the selection of drugs, schedule, and dosing for novel combination strategies., Competing Interests: Declaration of interests A.C.H. performed consulting work for Immunai and received research funding from Bristol Myers Squibb and Merck. R.S.H. has performed consulting work for Bristol Myers Squibb (exclusive of the current work). T.C.M. received honorarium for Scientific Advisory Board participation from: BMS, GigaGen, Merck, Pliant, Pfizer. G.C.K. is on the Merck Advisory Board. J.W. consulted for and have received less than $10,000 per annum from Merck, Genentech, AstraZeneca, GSK, Novartis, Nektar, Celldex, Incyte, Biond, Moderna, ImCheck, Sellas, Evaxion, Pfizer, Regeneron, and EMD Serono and received $10–$25,000 from BMS for membership on advisory boards. J.W. also holds equity in Biond, Evaxion, OncoC4, and Instil Bio, and on scientific advisory boards for CytomX, Incyte, ImCheck, Biond, Sellas, Instil Bio, OncoC4, and NexImmune and remunerated between $10,000–$50,000. In addition, J.W. is named on a patent filed by Moffitt Cancer Center on an ipilimumab biomarker and on TIL preparation and also on a PD-1 patent filed by Biodesix; J.W. receives less than $6000 in royalties. D.B., E.K., and C.S. were employed by Immunai when engaged in this project. S.G. and D.T. are employees of BMS. C.A. is a consultant for Biotherapy Partners., (Copyright © 2024 Elsevier Inc. All rights reserved.)- Published
- 2024
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3. Early Inflammation and Interferon Signaling Direct Enhanced Intestinal Crypt Regeneration after Proton FLASH Radiotherapy.
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Lim TL, Morral C, Verginadis II, Kim K, Luo L, Foley CJ, Kim MM, Li N, Yoshor B, Njah K, Putt M, Oliaei Motlagh SA, Velalopoulou A, Chowdhury P, Bicher S, Goia D, Lengner CJ, Wrana JL, Koumenis C, and Minn AJ
- Abstract
Ultra-high dose rate ("FLASH") radiotherapy (>40-60 Gy/s) is a promising new radiation modality currently in human clinical trials. Previous studies showed that FLASH proton radiotherapy (FR) improves toxicity of normal tissues compared to standard proton radiotherapy (SR) without compromising anti-tumor effects. Understanding this normal tissue sparing effect may offer insight into how toxicities from cancer therapy can be improved. Here, we show that compared to SR, FR resulted in improved acute weight recovery and survival in mice after whole-abdomen irradiation. Improved morbidity and mortality after FR were associated with greater proliferation of damage-induced epithelial progenitor cells followed by improved tissue regeneration. FR led to the accelerated differentiation of revival stem cells (revSCs), a rare damage-induced stem cell required for intestinal regeneration, and to qualitative and quantitative changes in activity of signaling pathways important for revSC differentiation and epithelial regeneration. Specifically, FR resulted in greater infiltration of macrophages producing TGF-β, a cytokine important for revSC induction, that was coupled to augmented TGF-β signaling in revSCs. In pericryptal fibroblasts, FR resulted in greater type I IFN (IFN-I) signaling, which directly stimulates production of FGF growth factors supporting revSC proliferation. Accordingly, the ability of FR to improve epithelial regeneration and morbidity was dependent on IFN-I signaling. In the context of SR, however, IFN-I had a detrimental effect and promoted toxicity. Thus, a tissue-level signaling network coordinated by differences in IFN-I signaling and involving stromal cells, immune cells, and revSCs underlies the ability of FLASH to improve normal tissue toxicity without compromising anti-tumor efficacy.
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- 2024
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4. Combined JAK inhibition and PD-1 immunotherapy for non-small cell lung cancer patients.
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Mathew D, Marmarelis ME, Foley C, Bauml JM, Ye D, Ghinnagow R, Ngiow SF, Klapholz M, Jun S, Zhang Z, Zorc R, Davis CW, Diehn M, Giles JR, Huang AC, Hwang WT, Zhang NR, Schoenfeld AJ, Carpenter EL, Langer CJ, Wherry EJ, and Minn AJ
- Subjects
- Animals, Female, Humans, Mice, Immunotherapy methods, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung immunology, Carcinoma, Non-Small-Cell Lung therapy, CD8-Positive T-Lymphocytes immunology, Immune Checkpoint Inhibitors therapeutic use, Janus Kinase 1 antagonists & inhibitors, Janus Kinase Inhibitors therapeutic use, Lung Neoplasms drug therapy, Lung Neoplasms immunology, Lung Neoplasms therapy, Programmed Cell Death 1 Receptor antagonists & inhibitors
- Abstract
Persistent inflammation driven by cytokines such as type-one interferon (IFN-I) can cause immunosuppression. We show that administration of the Janus kinase 1 (JAK1) inhibitor itacitinib after anti-PD-1 (programmed cell death protein 1) immunotherapy improves immune function and antitumor responses in mice and results in high response rates (67%) in a phase 2 clinical trial for metastatic non-small cell lung cancer. Patients who failed to respond to initial anti-PD-1 immunotherapy but responded after addition of itacitinib had multiple features of poor immune function to anti-PD-1 alone that improved after JAK inhibition. Itacitinib promoted CD8 T cell plasticity and therapeutic responses of exhausted and effector memory-like T cell clonotypes. Patients with persistent inflammation refractory to itacitinib showed progressive CD8 T cell terminal differentiation and progressive disease. Thus, JAK inhibition may improve the efficacy of anti-PD-1 immunotherapy by pivoting T cell differentiation dynamics.
- Published
- 2024
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5. p53 promotes revival stem cells in the regenerating intestine after severe radiation injury.
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Morral C, Ayyaz A, Kuo HC, Fink M, Verginadis II, Daniel AR, Burner DN, Driver LM, Satow S, Hasapis S, Ghinnagow R, Luo L, Ma Y, Attardi LD, Koumenis C, Minn AJ, Wrana JL, Lee CL, and Kirsch DG
- Subjects
- Mice, Animals, Intestines, Gastrointestinal Tract metabolism, Stem Cells metabolism, Apoptosis genetics, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, Radiation Injuries genetics, Radiation Injuries metabolism
- Abstract
Ionizing radiation induces cell death in the gastrointestinal (GI) epithelium by activating p53. However, p53 also prevents animal lethality caused by radiation-induced acute GI syndrome. Through single-cell RNA-sequencing of the irradiated mouse small intestine, we find that p53 target genes are specifically enriched in regenerating epithelial cells that undergo fetal-like reversion, including revival stem cells (revSCs) that promote animal survival after severe damage of the GI tract. Accordingly, in mice with p53 deleted specifically in the GI epithelium, ionizing radiation fails to induce fetal-like revSCs. Using intestinal organoids, we show that transient p53 expression is required for the induction of revival stem cells and is controlled by an Mdm2-mediated negative feedback loop. Together, our findings reveal that p53 suppresses severe radiation-induced GI injury by promoting fetal-like reprogramming of irradiated intestinal epithelial cells., (© 2024. The Author(s).)
- Published
- 2024
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6. Clinical and molecular features of acquired resistance to immunotherapy in non-small cell lung cancer.
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Memon D, Schoenfeld AJ, Ye D, Fromm G, Rizvi H, Zhang X, Keddar MR, Mathew D, Yoo KJ, Qiu J, Lihm J, Miriyala J, Sauter JL, Luo J, Chow A, Bhanot UK, McCarthy C, Vanderbilt CM, Liu C, Abu-Akeel M, Plodkowski AJ, McGranahan N, Łuksza M, Greenbaum BD, Merghoub T, Achour I, Barrett JC, Stewart R, Beltrao P, Schreiber TH, Minn AJ, Miller ML, and Hellmann MD
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- Humans, Animals, Mice, Signal Transduction, Immunotherapy, Antigen Presentation, B7-H1 Antigen metabolism, Tumor Microenvironment, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung genetics, Lung Neoplasms drug therapy, Lung Neoplasms genetics
- Abstract
Although immunotherapy with PD-(L)1 blockade is routine for lung cancer, little is known about acquired resistance. Among 1,201 patients with non-small cell lung cancer (NSCLC) treated with PD-(L)1 blockade, acquired resistance is common, occurring in >60% of initial responders. Acquired resistance shows differential expression of inflammation and interferon (IFN) signaling. Relapsed tumors can be separated by upregulated or stable expression of IFNγ response genes. Upregulation of IFNγ response genes is associated with putative routes of resistance characterized by signatures of persistent IFN signaling, immune dysfunction, and mutations in antigen presentation genes which can be recapitulated in multiple murine models of acquired resistance to PD-(L)1 blockade after in vitro IFNγ treatment. Acquired resistance to PD-(L)1 blockade in NSCLC is associated with an ongoing, but altered IFN response. The persistently inflamed, rather than excluded or deserted, tumor microenvironment of acquired resistance may inform therapeutic strategies to effectively reprogram and reverse acquired resistance., Competing Interests: Declaration of interests A.J.S. reports consulting/advising role to J&J, KSQ therapeutics, BMS, Merck, Enara Bio, Perceptive Advisors, Oppenheimer and Co, Umoja Biopharma, Legend Biotech, Iovance Biotherapeutics, Prelude Therapeutics, Immunocore, Lyell Immunopharma, Amgen and Heat Biologics. Research funding: GSK (Inst), PACT pharma (Inst), Iovance Biotherapeutics (Inst), Achilles therapeutics (Inst), Merck (Inst), BMS (Inst), Harpoon Therapeutics (Inst) and Amgen (Inst). MDH reports research grant from BMS; personal fees from Achilles; Arcus; AstraZeneca; Blueprint; BMS; Genentech/Roche; Genzyme/Sanofi, Immunai; Instil Bio; Janssen; Merck; Mirati; Natera; Nektar; Pact Pharma; Regeneron; Shattuck Labs; Syndax; as well as equity options from Arcus, Factorial, Immunai, and Shattuck Labs. A patent filed by Memorial Sloan Kettering related to the use of tumor mutational burden to predict response to immunotherapy (PCT/US2015/062208) is pending and licensed by PGDx. J.L. has received honoraria from Targeted Oncology and Physicians’ Education Resource. D.M. is an employee of M:M Bio Limited. D.M. reports consulting role to Shattuck Labs and Corbus Pharma. T.M. is a consultant for Daiichi Sankyo Co, Leap Therapeutics, Immunos Therapeutics, and Pfizer, and co-founder of Imvaq Therapeutics. T.M. has equity in Imvaq therapeutics. T.M. reports grants from Bristol Myers Squibb, Surface Oncology, Kyn Therapeutics, Infinity Pharmaceuticals, Peregrine Pharmaceuticals, Adaptive Biotechnologies, Leap Therapeutics, and Aprea. T.M. is an inventor on patent applications related to work on oncolytic viral therapy, alphavirus-based vaccines, neo-antigen modeling, CD40, GITR, OX40, PD-1, and CTLA-4. B.D.G. has received honoraria for speaking engagements from Merck, Bristol Meyers Squibb, and Chugai Pharmaceuticals; has received research funding from Bristol Meyers Squibb and Merck; and has been a compensated consultant for Darwin Health, Merck, PMV Pharma, Shennon Biotechnologies, and Rome Therapeutics of which he is a co-founder. B.D.G. is part of a patent related to neoantigen prediction (WO2018136664A1, PCT/US2023/011643). G.F. and T.H.S. are employees and stockholders of Shattuck Labs, Inc. M.L.M. has received honorarium from GSK. H.R., X.Z., M.R.K., I.A., R.S., J.C.B., M.L.M., and M.D.H. are current employees and stockholders of AstraZeneca., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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7. Signal recovery in single cell batch integration.
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Zhang Z, Mathew D, Lim T, Mason K, Martinez CM, Huang S, Wherry EJ, Susztak K, Minn AJ, Ma Z, and Zhang NR
- Abstract
Data integration to align cells across batches has become a cornerstone of single cell data analysis, critically affecting downstream results. Yet, how much biological signal is erased during integration? Currently, there are no guidelines for when the biological differences between samples are separable from batch effects, and thus, data integration usually involve a lot of guesswork: Cells across batches should be aligned to be "appropriately" mixed, while preserving "main cell type clusters". We show evidence that current paradigms for single cell data integration are unnecessarily aggressive, removing biologically meaningful variation. To remedy this, we present a novel statistical model and computationally scalable algorithm, CellANOVA, to recover biological signal that is lost during single cell data integration. CellANOVA utilizes a "pool-of-controls" design concept, applicable across diverse settings, to separate unwanted variation from biological variation of interest. When applied with existing integration methods, CellANOVA allows the recovery of subtle biological signals and corrects, to a large extent, the data distortion introduced by integration. Further, CellANOVA explicitly estimates cell- and gene-specific batch effect terms which can be used to identify the cell types and pathways exhibiting the largest batch variations, providing clarity as to which biological signals can be recovered. These concepts are illustrated on studies of diverse designs, where the biological signals that are recovered by CellANOVA are shown to be validated by orthogonal assays. In particular, we show that CellANOVA is effective in the challenging case of single-cell and single-nuclei data integration, where the recovered biological signals are replicated in an independent study.
- Published
- 2023
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8. Isolation of Epithelial and Stromal Cells from Colon Tissues in Homeostasis and Under Inflammatory Conditions.
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Morral C, Ghinnagow R, Karakasheva T, Zhou Y, Thadi A, Li N, Yoshor B, Soto GE, Chen CH, Aleynick D, Weinbrom S, Fulton M, Uzun Y, Bewtra M, Kelsen JR, Lengner CJ, Tan K, Minn AJ, and Hamilton KE
- Abstract
Inflammation of the gastrointestinal tract is a prevalent pathology in diseases such as inflammatory bowel disease (IBD). Currently, there are no therapies to prevent IBD, and available therapies to treat IBD are often sub-optimal. Thus, an unmet need exists to better understand the molecular mechanisms underlying intestinal tissue responses to damage and regeneration. The recent development of single-cell RNA (sc-RNA) sequencing-based techniques offers a unique opportunity to shed light on novel signaling pathways and cellular states that govern tissue adaptation or maladaptation across a broad spectrum of diseases. These approaches require the isolation of high-quality cells from tissues for downstream transcriptomic analyses. In the context of intestinal biology, there is a lack of protocols that ensure the isolation of epithelial and non-epithelial compartments simultaneously with high-quality yield. Here, we report two protocols for the isolation of epithelial and stromal cells from mouse and human colon tissues under inflammatory conditions. Specifically, we tested the feasibility of the protocols in a mouse model of dextran sodium sulfate (DSS)-induced colitis and in human biopsies from Crohn's patients. We performed sc-RNA sequencing analysis and demonstrated that the protocol preserves most of the epithelial and stromal cell types found in the colon. Moreover, the protocol is suitable for immunofluorescence staining of surface markers for epithelial, stromal, and immune cell lineages for flow cytometry analyses. This optimized protocol will provide a new resource for scientists to study complex tissues such as the colon in the context of tissue damage and regeneration. Key features • This protocol allows the isolation of epithelial and stromal cells from colon tissues. • The protocol has been optimized for tissues under inflammatory conditions with compromised cell viability. • This protocol is suitable for experimental mouse models of colon inflammation and human biopsies., (©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.)
- Published
- 2023
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9. RUNX1 is required in granulocyte-monocyte progenitors to attenuate inflammatory cytokine production by neutrophils.
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Zezulin AU, Yen D, Ye D, Howell ED, Bresciani E, Diemer J, Ren JG, Ahmad MH, Castilla LH, Touw IP, Minn AJ, Tong W, Liu PP, Tan K, Yu W, and Speck NA
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- Monocytes metabolism, Core Binding Factor Alpha 2 Subunit genetics, Core Binding Factor Alpha 2 Subunit metabolism, Cytokines metabolism, Chromatin metabolism, STAT1 Transcription Factor metabolism, Neutrophils, Toll-Like Receptor 4 metabolism
- Abstract
The transcription factor RUNX1 is mutated in familial platelet disorder with associated myeloid malignancy (FPDMM) and in sporadic myelodysplastic syndrome and leukemia. RUNX1 was shown to regulate inflammation in multiple cell types. Here we show that RUNX1 is required in granulocyte-monocyte progenitors (GMPs) to epigenetically repress two inflammatory signaling pathways in neutrophils: Toll-like receptor 4 (TLR4) and type I interferon (IFN) signaling. RUNX1 loss in GMPs augments neutrophils' inflammatory response to the TLR4 ligand lipopolysaccharide through increased expression of the TLR4 coreceptor CD14. RUNX1 binds Cd14 and other genes encoding proteins in the TLR4 and type I IFN signaling pathways whose chromatin accessibility increases when RUNX1 is deleted. Transcription factor footprints for the effectors of type I IFN signaling-the signal transducer and activator of transcription (STAT1::STAT2) and interferon regulatory factors (IRFs)-were enriched in chromatin that gained accessibility in both GMPs and neutrophils when RUNX1 was lost. STAT1::STAT2 and IRF motifs were also enriched in the chromatin of retrotransposons that were derepressed in RUNX1-deficient GMPs and neutrophils. We conclude that a major direct effect of RUNX1 loss in GMPs is the derepression of type I IFN and TLR4 signaling, resulting in a state of fixed maladaptive innate immunity., (© 2023 Zezulin et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2023
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10. TLR priming licenses NAIP inflammasome activation by immunoevasive ligands.
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Grayczyk JP, Egan MS, Liu L, Aunins E, Wynosky-Dolfi MA, Canna S, Minn AJ, Shin S, and Brodsky IE
- Abstract
NLR family, apoptosis inhibitory proteins (NAIPs) detect bacterial flagellin and structurally related components of bacterial type III secretion systems (T3SS), and recruit NLR family, CARD domain containing protein 4 (NLRC4) and caspase-1 into an inflammasome complex that induces pyroptosis. NAIP/NLRC4 inflammasome assembly is initiated by the binding of a single NAIP to its cognate ligand, but a subset of bacterial flagellins or T3SS structural proteins are thought to evade NAIP/NLRC4 inflammasome sensing by not binding to their cognate NAIPs. Unlike other inflammasome components such as NLRP3, AIM2, or some NAIPs, NLRC4 is constitutively present in resting macrophages, and not thought to be regulated by inflammatory signals. Here, we demonstrate that Toll-like receptor (TLR) stimulation upregulates NLRC4 transcription and protein expression in murine macrophages, which licenses NAIP detection of evasive ligands. TLR-induced NLRC4 upregulation and NAIP detection of evasive ligands required p38 MAPK signaling. In contrast, TLR priming in human macrophages did not upregulate NLRC4 expression, and human macrophages remained unable to detect NAIP-evasive ligands even following priming. Critically, ectopic expression of either murine or human NLRC4 was sufficient to induce pyroptosis in response to immunoevasive NAIP ligands, indicating that increased levels of NLRC4 enable the NAIP/NLRC4 inflammasome to detect these normally evasive ligands. Altogether, our data reveal that TLR priming tunes the threshold for NAIP/NLRC4 inflammasome activation and enables inflammasome responses against immunoevasive or suboptimal NAIP ligands.
- Published
- 2023
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11. p53 promotes revival stem cells in the regenerating intestine after severe radiation injury.
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Morral C, Ayyaz A, Kuo HC, Fink M, Verginadis I, Daniel AR, Burner DN, Driver LM, Satow S, Hasapis S, Ghinnagow R, Luo L, Ma Y, Attardi LD, Koumenis C, Minn AJ, Wrana JL, Lee CL, and Kirsch DG
- Abstract
Ionizing radiation induces cell death in the gastrointestinal (GI) epithelium by activating p53. However, p53 also prevents animal lethality caused by radiation-induced GI injury. Through single-cell RNA-sequencing of the irradiated mouse intestine, we find that p53 target genes are specifically enriched in stem cells of the regenerating epithelium, including revival stem cells that promote animal survival after GI damage. Accordingly, in mice with p53 deleted specifically in the GI epithelium, ionizing radiation fails to induce revival stem cells. Using intestinal organoids, we show that transient p53 expression is required for the induction of revival stem cells that is controlled by an Mdm2-mediated negative feedback loop. These results suggest that p53 suppresses severe radiation-indued GI injury by promoting intestinal epithelial cell reprogramming., One-Sentence Summary: After severe radiation injury to the intestine, transient p53 activity induces revival stem cells to promote regeneration.
- Published
- 2023
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12. Cancer cells resistant to immune checkpoint blockade acquire interferon-associated epigenetic memory to sustain T cell dysfunction.
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Qiu J, Xu B, Ye D, Ren D, Wang S, Benci JL, Xu Y, Ishwaran H, Beltra JC, Wherry EJ, Shi J, and Minn AJ
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- Animals, Humans, Mice, Epigenetic Memory, Immune Checkpoint Inhibitors pharmacology, Immune Checkpoint Inhibitors therapeutic use, Interferon-gamma genetics, Interferon-gamma metabolism, Interferon-gamma pharmacology, Signal Transduction, T-Lymphocytes immunology, Interferon Type I metabolism, Interferon Type I pharmacology, Neoplasms drug therapy, Neoplasms genetics
- Abstract
Prolonged interferon (IFN) signaling in cancer cells can promote resistance to immune checkpoint blockade (ICB). How cancer cells retain effects of prolonged IFN stimulation to coordinate resistance is unclear. We show that, across human and/or mouse tumors, immune dysfunction is associated with cancer cells acquiring epigenetic features of inflammatory memory. Here, inflammatory memory domains, many of which are initiated by chronic IFN-γ, are maintained by signal transducer and activator of transcription (STAT)1 and IFN regulatory factor (IRF)3 and link histone 3 lysine 4 monomethylation (H3K4me1)-marked chromatin accessibility to increased expression of a subset of IFN-stimulated genes (ISGs). These ISGs include the RNA sensor OAS1 that amplifies type I IFN (IFN-I) and immune inhibitory genes. Abrogating cancer cell IFN-I signaling restores anti-programmed cell death protein 1 (PD1) response by increasing IFN-γ in immune cells, promoting dendritic cell and CD8
+ T cell interactions, and expanding T cells toward effector-like states rather than exhausted states. Thus, cancer cells acquire inflammatory memory to augment a subset of ISGs that promote and predict IFN-driven immune dysfunction., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)- Published
- 2023
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13. Targeting PARP11 to avert immunosuppression and improve CAR T therapy in solid tumors.
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Zhang H, Yu P, Tomar VS, Chen X, Atherton MJ, Lu Z, Zhang HG, Li S, Ortiz A, Gui J, Leu NA, Yan F, Blanco A, Meyer-Ficca ML, Meyer RG, Beiting DP, Li J, Nunez-Cruz S, O'Connor RS, Johnson LR, Minn AJ, George SS, Koumenis C, Diehl JA, Milone MC, Zheng H, and Fuchs SY
- Subjects
- Humans, Immunosuppression Therapy, Immunotherapy, Adoptive, Tumor Microenvironment, Neoplasms drug therapy, Poly(ADP-ribose) Polymerases metabolism, Receptors, Chimeric Antigen genetics
- Abstract
Evasion of antitumor immunity and resistance to therapies in solid tumors are aided by an immunosuppressive tumor microenvironment (TME). We found that TME factors, such as regulatory T cells and adenosine, downregulated type I interferon receptor IFNAR1 on CD8
+ cytotoxic T lymphocytes (CTLs). These events relied upon poly-ADP ribose polymerase-11 (PARP11), which was induced in intratumoral CTLs and acted as a key regulator of the immunosuppressive TME. Ablation of PARP11 prevented loss of IFNAR1, increased CTL tumoricidal activity and inhibited tumor growth in an IFNAR1-dependent manner. Accordingly, genetic or pharmacologic inactivation of PARP11 augmented the therapeutic benefits of chimeric antigen receptor T cells. Chimeric antigen receptor CTLs engineered to inactivate PARP11 demonstrated a superior efficacy against solid tumors. These findings highlight the role of PARP11 in the immunosuppressive TME and provide a proof of principle for targeting this pathway to optimize immune therapies., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)- Published
- 2022
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14. The interferon-stimulated gene RIPK1 regulates cancer cell intrinsic and extrinsic resistance to immune checkpoint blockade.
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Cucolo L, Chen Q, Qiu J, Yu Y, Klapholz M, Budinich KA, Zhang Z, Shao Y, Brodsky IE, Jordan MS, Gilliland DG, Zhang NR, Shi J, and Minn AJ
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- Animals, Immunotherapy, Interferon-gamma metabolism, Mice, NF-kappa B metabolism, Drug Resistance, Neoplasm, Immune Checkpoint Inhibitors, Interferons metabolism, Neoplasms genetics, Receptor-Interacting Protein Serine-Threonine Kinases genetics, Receptor-Interacting Protein Serine-Threonine Kinases metabolism
- Abstract
Interferon-gamma (IFN-γ) has pleiotropic effects on cancer immune checkpoint blockade (ICB), including roles in ICB resistance. We analyzed gene expression in ICB-sensitive versus ICB-resistant tumor cells and identified a strong association between interferon-mediated resistance and expression of Ripk1, a regulator of tumor necrosis factor (TNF) superfamily receptors. Genetic interaction screening revealed that in cancer cells, RIPK1 diverted TNF signaling through NF-κB and away from its role in cell death. This promoted an immunosuppressive chemokine program by cancer cells, enhanced cancer cell survival, and decreased infiltration of T and NK cells expressing TNF superfamily ligands. Deletion of RIPK1 in cancer cells compromised chemokine secretion, decreased ARG1
+ suppressive myeloid cells linked to ICB failure in mice and humans, and improved ICB response driven by CASP8-killing and dependent on T and NK cells. RIPK1-mediated resistance required its ubiquitin scaffolding but not kinase function. Thus, cancer cells co-opt RIPK1 to promote cell-intrinsic and cell-extrinsic resistance to immunotherapy., Competing Interests: Declaration of interests A.J.M. has received research funding from Merck. He is a scientific advisor for Takeda, H3Biomedicine, Xilio, and Related Sciences. A.J.M. is an inventor on patents related to the IFN pathway and an inventor on a filed patent related to modified CAR T cells. A.J.M. is a scientific founder for Dispatch Biotherapeutics., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
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15. The immunostimulatory RNA RN7SL1 enables CAR-T cells to enhance autonomous and endogenous immune function.
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Johnson LR, Lee DY, Eacret JS, Ye D, June CH, and Minn AJ
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- Animals, Antigens metabolism, CD8-Positive T-Lymphocytes immunology, Cell Line, Tumor, DEAD Box Protein 58 metabolism, Dendritic Cells drug effects, Dendritic Cells metabolism, Extracellular Vesicles metabolism, Humans, Immunity drug effects, Immunocompetence, Immunologic Memory, Immunotherapy, Interferons metabolism, Melanoma, Experimental pathology, Mice, Inbred C57BL, Myeloid Cells drug effects, Myeloid Cells metabolism, Peptides metabolism, Receptors, Pattern Recognition metabolism, T-Lymphocytes drug effects, Mice, Immunologic Factors pharmacology, RNA pharmacology, Receptors, Chimeric Antigen immunology, T-Lymphocytes immunology
- Abstract
Poor tumor infiltration, development of exhaustion, and antigen insufficiency are common mechanisms that limit chimeric antigen receptor (CAR)-T cell efficacy. Delivery of pattern recognition receptor agonists is one strategy to improve immune function; however, targeting these agonists to immune cells is challenging, and off-target signaling in cancer cells can be detrimental. Here, we engineer CAR-T cells to deliver RN7SL1, an endogenous RNA that activates RIG-I/MDA5 signaling. RN7SL1 promotes expansion and effector-memory differentiation of CAR-T cells. Moreover, RN7SL1 is deployed in extracellular vesicles and selectively transferred to immune cells. Unlike other RNA agonists, transferred RN7SL1 restricts myeloid-derived suppressor cell (MDSC) development, decreases TGFB in myeloid cells, and fosters dendritic cell (DC) subsets with costimulatory features. Consequently, endogenous effector-memory and tumor-specific T cells also expand, allowing rejection of solid tumors with CAR antigen loss. Supported by improved endogenous immunity, CAR-T cells can now co-deploy peptide antigens with RN7SL1 to enhance efficacy, even when heterogenous CAR antigen tumors lack adequate neoantigens., Competing Interests: Declaration of interests A.J.M. has received research funding from Merck. He is a scientific advisor for Takeda, H3Biomedicine, Related Sciences, and Xilio. A.J.M. is an inventor on patents related to the IFN pathway. A.J.M., L.R.J., and C.H.J. are inventors on a filed patent related to modified CAR-T cells. C.H.J. reports research funding from Novartis, and he is a scientific founder of Tmunity Therapeutics. A.J.M., C.H.J., and L.R.J. are scientific founders for Project 5 Therapeutics. C.H.J. also works under a research collaboration involving the University of Pennsylvania and the Novartis Institute of Biomedical Research and is an inventor of intellectual property licensed by the University of Pennsylvania to Novartis. C.H.J. is on the board of directors for AC Immune and is a scientific advisor for BluesphereBio, Cabaletta, Carisma, Cartography, Cellares, Celldex, DeCART, Decheng, Poseida, Verismo, WIRB Copernicus, and Ziopharm., (Copyright © 2021. Published by Elsevier Inc.)
- Published
- 2021
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16. FLASH Proton Radiotherapy Spares Normal Epithelial and Mesenchymal Tissues While Preserving Sarcoma Response.
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Velalopoulou A, Karagounis IV, Cramer GM, Kim MM, Skoufos G, Goia D, Hagan S, Verginadis II, Shoniyozov K, Chiango J, Cerullo M, Varner K, Yao L, Qin L, Hatzigeorgiou AG, Minn AJ, Putt M, Lanza M, Assenmacher CA, Radaelli E, Huck J, Diffenderfer E, Dong L, Metz J, Koumenis C, Cengel KA, Maity A, and Busch TM
- Subjects
- Animals, Bone and Bones pathology, Bone and Bones radiation effects, Disease Models, Animal, Dogs, Female, Gene Expression Profiling, Humans, Mice, Morbidity, Muscles pathology, Muscles radiation effects, Radiation Injuries diagnosis, Radiation Injuries etiology, Radiotherapy Dosage, Sarcoma metabolism, Skin radiation effects, Treatment Outcome, Epithelium radiation effects, Organ Sparing Treatments methods, Proton Therapy adverse effects, Proton Therapy methods, Sarcoma pathology, Sarcoma radiotherapy
- Abstract
In studies of electron and proton radiotherapy, ultrahigh dose rates of FLASH radiotherapy appear to produce fewer toxicities than standard dose rates while maintaining local tumor control. FLASH-proton radiotherapy (F-PRT) brings the spatial advantages of PRT to FLASH dose rates (>40 Gy/second), making it important to understand if and how F-PRT spares normal tissues while providing antitumor efficacy that is equivalent to standard-proton radiotherapy (S-PRT). Here we studied PRT damage to skin and mesenchymal tissues of muscle and bone and found that F-PRT of the C57BL/6 murine hind leg produced fewer severe toxicities leading to death or requiring euthanasia than S-PRT of the same dose. RNA-seq analyses of murine skin and bone revealed pathways upregulated by S-PRT yet unaltered by F-PRT, such as apoptosis signaling and keratinocyte differentiation in skin, as well as osteoclast differentiation and chondrocyte development in bone. Corroborating these findings, F-PRT reduced skin injury, stem cell depletion, and inflammation, mitigated late effects including lymphedema, and decreased histopathologically detected myofiber atrophy, bone resorption, hair follicle atrophy, and epidermal hyperplasia. F-PRT was equipotent to S-PRT in control of two murine sarcoma models, including at an orthotopic intramuscular site, thereby establishing its relevance to mesenchymal cancers. Finally, S-PRT produced greater increases in TGFβ1 in murine skin and the skin of canines enrolled in a phase I study of F-PRT versus S-PRT. Collectively, these data provide novel insights into F-PRT-mediated tissue sparing and support its ongoing investigation in applications that would benefit from this sparing of skin and mesenchymal tissues. SIGNIFICANCE: These findings will spur investigation of FLASH radiotherapy in sarcoma and additional cancers where mesenchymal tissues are at risk, including head and neck cancer, breast cancer, and pelvic malignancies., (©2021 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2021
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17. Thymic stromal lymphopoietin induces adipose loss through sebum hypersecretion.
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Choa R, Tohyama J, Wada S, Meng H, Hu J, Okumura M, May RM, Robertson TF, Pai RL, Nace A, Hopkins C, Jacobsen EA, Haldar M, FitzGerald GA, Behrens EM, Minn AJ, Seale P, Cotsarelis G, Kim B, Seykora JT, Li M, Arany Z, and Kambayashi T
- Subjects
- Adaptive Immunity, Animals, Cytokines genetics, Diet, Glucose metabolism, Homeostasis, Humans, Immunoglobulins metabolism, Lipid Metabolism, Mice, Non-alcoholic Fatty Liver Disease prevention & control, Obesity prevention & control, Pore Forming Cytotoxic Proteins metabolism, Receptors, Cytokine metabolism, Sebaceous Glands metabolism, Signal Transduction, Skin immunology, T-Lymphocytes physiology, Weight Loss, Thymic Stromal Lymphopoietin, Adipose Tissue, White anatomy & histology, Cytokines metabolism, Sebum metabolism, Skin metabolism
- Abstract
Emerging studies indicate that the immune system can regulate systemic metabolism. Here, we show that thymic stromal lymphopoietin (TSLP) stimulates T cells to induce selective white adipose loss, which protects against obesity, improves glucose metabolism, and mitigates nonalcoholic steatohepatitis. Unexpectedly, adipose loss was not caused by alterations in food intake, absorption, or energy expenditure. Rather, it was induced by the excessive loss of lipids through the skin as sebum. TSLP and T cells regulated sebum release and sebum-associated antimicrobial peptide expression in the steady state. In human skin, TSLP expression correlated directly with sebum-associated gene expression. Thus, we establish a paradigm in which adipose loss can be achieved by means of sebum hypersecretion and uncover a role for adaptive immunity in skin barrier function through sebum secretion., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2021
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18. A stratified phase I dose escalation trial of hypofractionated radiotherapy followed by ipilimumab in metastatic melanoma: long-term follow-up and final outcomes.
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Maity A, Mick R, Rengan R, Mitchell TC, Amaravadi RK, Schuchter LM, Pryma DA, Patsch DM, Maity AP, Minn AJ, Vonderheide RH, and Lukens JN
- Subjects
- Follow-Up Studies, Humans, Ipilimumab therapeutic use, Progression-Free Survival, Melanoma drug therapy, Neoplasms, Second Primary
- Abstract
We conducted a phase I dose-escalation trial of radiation with ipilimumab in patients with melanoma with ≥2 metastatic lesions. Here, we report the final full clinical analysis. Patients received RT (6 or 8 Gy x 2 or 3 doses) to a single lesion followed by 4 cycles of ipilimumab. The primary endpoint was maximum tolerated dose of RT, and secondary endpoint was response at non-radiated sites. Twenty-two patients with treatment-naïve (n = 11) or treatment-refractory (n = 11) Stage IV melanoma were enrolled. There were 31 treatment-related adverse events (AEs), of which 16 were deemed immune-related. Eleven patients had grade 3 AEs (no grade 4/5). There were no dose-limiting toxicities related to the radiation/ipilimumab combination. Five of 22 patients (22.7%, 95% CI 7.8-45.4%) had partial response as best response and three (13.6%) had stable disease. Median overall survival was 10.7 months (95% CI, 4.9 months to not-estimable) and median progression-free survival 3.6 months (95% CI, 2.9 months to 7.8 months). Seven patients were still alive at the time of last follow-up (median follow-up 89.2 months), most of whom received pembrolizumab after progression. Radiotherapy followed by ipilimumab was well tolerated and yielded a response rate that compares favorably to the objective response rate with ipilimumab alone. Furthermore, 32% of patients are long-term survivors, most of whom received pembrolizumab. Based on these results, the recommended dose that was used in subsequent Phase 2 trials was 8 Gy x 3 doses. Clinical Trial Registration: NCT01497808 (www.clinicaltrials.gov)., (© 2021 The Author(s). Published with license by Taylor & Francis Group, LLC.)
- Published
- 2021
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19. Cell Cycle Checkpoints Cooperate to Suppress DNA- and RNA-Associated Molecular Pattern Recognition and Anti-Tumor Immune Responses.
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Chen J, Harding SM, Natesan R, Tian L, Benci JL, Li W, Minn AJ, Asangani IA, and Greenberg RA
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- Humans, Neoplasms pathology, Signal Transduction, Cell Cycle Proteins metabolism, DNA genetics, Immunity genetics, Neoplasms immunology, RNA genetics
- Abstract
The DNA-dependent pattern recognition receptor, cGAS (cyclic GMP-AMP synthase), mediates communication between the DNA damage and the immune responses. Mitotic chromosome missegregation stimulates cGAS activity; however, it is unclear whether progression through mitosis is required for cancercell-intrinsic activation of anti-tumor immune responses. Moreover, it is unknown whether cell cycle checkpoint disruption can restore responses in cancer cells that are recalcitrant to DNAdamage-induced inflammation. Here, we demonstrate that prolonged cell cycle arrest at the G
2 -mitosis boundary from either excessive DNA damage or CDK1 inhibition prevents inflammatory-stimulated gene expression and immune-mediated destruction of distal tumors. Remarkably, DNAdamage-induced inflammatory signaling is restored in a RIG-I-dependent manner upon concomitant disruption of p53 and the G2 checkpoint. These findings link aberrant cell progression and p53 loss to an expanded spectrum of damage-associated molecular pattern recognition and have implications for the design of rational approaches to augment anti-tumor immune responses., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2020
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20. Combining Radiation with Immunotherapy: The University of Pennsylvania Experience.
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Luo LY, O'Hara MH, Mitchell TC, Vonderheide RH, Wherry EJ, Minn AJ, and Maity A
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- Animals, Antibodies, Monoclonal therapeutic use, Antigens, Neoplasm immunology, Clinical Trials as Topic, Combined Modality Therapy, Humans, Immunotherapy methods, Interferons immunology, Pennsylvania, Research Design, Signal Transduction drug effects, Signal Transduction radiation effects, Immune Checkpoint Inhibitors therapeutic use, Neoplasms immunology, Neoplasms radiotherapy
- Abstract
Immune checkpoint inhibitors have shown remarkable clinical benefit across a variety of cancer types. However, the majority of patients do not respond or develop relapse after therapy. Radiation can favorably modulate the immune system and enhance tumor antigen recognition and rejection. Thus, the combination of radiation and immune checkpoint blockade (ICB) has been recognized as a promising strategy to improve tumor response and broaden the clinical utility of immunotherapy. In this review, we highlight the preclinical and clinical experience at our institution aimed at understanding and promoting the immunostimulatory effect of radiation. We discuss the rationale, design, results, and lessons from our clinical trials in combining radiation with anti-CTLA4 and/or anti-PD-1 therapy. In parallel, our studies to understand the resistance mechanism to radiation and ICB have converged on interferon (IFN) signaling as a key regulatory pathway. Persistent IFN-γ signaling impairs anti-tumor immune responses which can be reversed by using JAK inhibitor to disrupt the IFN signaling. Lastly we discuss remaining challenges, ongoing studies, and future directions in combining radiation with immunotherapy., (Copyright © 2020 Elsevier Inc. All rights reserved.)
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- 2020
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21. Opposing Functions of Interferon Coordinate Adaptive and Innate Immune Responses to Cancer Immune Checkpoint Blockade.
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Benci JL, Johnson LR, Choa R, Xu Y, Qiu J, Zhou Z, Xu B, Ye D, Nathanson KL, June CH, Wherry EJ, Zhang NR, Ishwaran H, Hellmann MD, Wolchok JD, Kambayashi T, and Minn AJ
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- Adoptive Transfer, Animals, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, CD8-Positive T-Lymphocytes immunology, CTLA-4 Antigen antagonists & inhibitors, Cell Line, Tumor, Cohort Studies, Female, Gene Knockout Techniques, Humans, Interferon-gamma antagonists & inhibitors, Killer Cells, Natural immunology, Lung Neoplasms drug therapy, Melanoma drug therapy, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, Programmed Cell Death 1 Receptor antagonists & inhibitors, Progression-Free Survival, RNA-Seq, Transfection, Adaptive Immunity immunology, Immunity, Innate immunology, Interferon-gamma genetics, Interferon-gamma metabolism, Lung Neoplasms immunology, Melanoma immunology
- Abstract
Interferon-gamma (IFNG) augments immune function yet promotes T cell exhaustion through PDL1. How these opposing effects are integrated to impact immune checkpoint blockade (ICB) is unclear. We show that while inhibiting tumor IFNG signaling decreases interferon-stimulated genes (ISGs) in cancer cells, it increases ISGs in immune cells by enhancing IFNG produced by exhausted T cells (T
EX ). In tumors with favorable antigenicity, these TEX mediate rejection. In tumors with neoantigen or MHC-I loss, TEX instead utilize IFNG to drive maturation of innate immune cells, including a PD1+ TRAIL+ ILC1 population. By disabling an inhibitory circuit impacting PD1 and TRAIL, blocking tumor IFNG signaling promotes innate immune killing. Thus, interferon signaling in cancer cells and immune cells oppose each other to establish a regulatory relationship that limits both adaptive and innate immune killing. In melanoma and lung cancer patients, perturbation of this relationship is associated with ICB response independent of tumor mutational burden., (Copyright © 2019 Elsevier Inc. All rights reserved.)- Published
- 2019
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22. Correction to: Toward a comprehensive view of cancer immune responsiveness: a synopsis from the SITC workshop.
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Bedognetti D, Ceccarelli M, Galluzzi L, Lu R, Palucka K, Samayoa J, Spranger S, Warren S, Wong KK, Ziv E, Chowell D, Coussens LM, De Carvalho DD, DeNardo DG, Galon J, Kaufman HL, Kirchhoff T, Lotze MT, Luke JJ, Minn AJ, Politi K, Shultz LD, Simon R, Thórsson V, Weidhaas JB, Ascierto ML, Ascierto PA, Barnes JM, Barsan V, Bommareddy PK, Bot A, Church SE, Ciliberto G, De Maria A, Draganov D, Ho WS, McGee HM, Monette A, Murphy JF, Nisticò P, Park W, Patel M, Quigley M, Radvanyi L, Raftopoulos H, Rudqvist NP, Snyder A, Sweis RF, Valpione S, Zappasodi R, Butterfield LH, Disis ML, Fox BA, Cesano A, and Marincola FM
- Abstract
Following publication of the original article [1], the author reported that an author name, Roberta Zappasodi, was missed in the authorship list.
- Published
- 2019
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23. Toward a comprehensive view of cancer immune responsiveness: a synopsis from the SITC workshop.
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Bedognetti D, Ceccarelli M, Galluzzi L, Lu R, Palucka K, Samayoa J, Spranger S, Warren S, Wong KK, Ziv E, Chowell D, Coussens LM, De Carvalho DD, DeNardo DG, Galon J, Kaufman HL, Kirchhoff T, Lotze MT, Luke JJ, Minn AJ, Politi K, Shultz LD, Simon R, Thórsson V, Weidhaas JB, Ascierto ML, Ascierto PA, Barnes JM, Barsan V, Bommareddy PK, Bot A, Church SE, Ciliberto G, De Maria A, Draganov D, Ho WS, McGee HM, Monette A, Murphy JF, Nisticò P, Park W, Patel M, Quigley M, Radvanyi L, Raftopoulos H, Rudqvist NP, Snyder A, Sweis RF, Valpione S, Zappasodi R, Butterfield LH, Disis ML, Fox BA, Cesano A, and Marincola FM
- Subjects
- Advisory Committees, Animals, Biomarkers, Tumor genetics, Biomarkers, Tumor immunology, Congresses as Topic, Disease Models, Animal, Humans, Medical Oncology organization & administration, Neoplasms genetics, Neoplasms immunology, Societies, Medical organization & administration, Treatment Outcome, Tumor Microenvironment genetics, Immunotherapy, Neoplasms therapy, Tumor Microenvironment immunology
- Abstract
Tumor immunology has changed the landscape of cancer treatment. Yet, not all patients benefit as cancer immune responsiveness (CIR) remains a limitation in a considerable proportion of cases. The multifactorial determinants of CIR include the genetic makeup of the patient, the genomic instability central to cancer development, the evolutionary emergence of cancer phenotypes under the influence of immune editing, and external modifiers such as demographics, environment, treatment potency, co-morbidities and cancer-independent alterations including immune homeostasis and polymorphisms in the major and minor histocompatibility molecules, cytokines, and chemokines. Based on the premise that cancer is fundamentally a disorder of the genes arising within a cell biologic process, whose deviations from normality determine the rules of engagement with the host's response, the Society for Immunotherapy of Cancer (SITC) convened a task force of experts from various disciplines including, immunology, oncology, biophysics, structural biology, molecular and cellular biology, genetics, and bioinformatics to address the complexity of CIR from a holistic view. The task force was launched by a workshop held in San Francisco on May 14-15, 2018 aimed at two preeminent goals: 1) to identify the fundamental questions related to CIR and 2) to create an interactive community of experts that could guide scientific and research priorities by forming a logical progression supported by multiple perspectives to uncover mechanisms of CIR. This workshop was a first step toward a second meeting where the focus would be to address the actionability of some of the questions identified by working groups. In this event, five working groups aimed at defining a path to test hypotheses according to their relevance to human cancer and identifying experimental models closest to human biology, which include: 1) Germline-Genetic, 2) Somatic-Genetic and 3) Genomic-Transcriptional contributions to CIR, 4) Determinant(s) of Immunogenic Cell Death that modulate CIR, and 5) Experimental Models that best represent CIR and its conversion to an immune responsive state. This manuscript summarizes the contributions from each group and should be considered as a first milestone in the path toward a more contemporary understanding of CIR. We appreciate that this effort is far from comprehensive and that other relevant aspects related to CIR such as the microbiome, the individual's recombined T cell and B cell receptors, and the metabolic status of cancer and immune cells were not fully included. These and other important factors will be included in future activities of the taskforce. The taskforce will focus on prioritization and specific actionable approach to answer the identified questions and implementing the collaborations in the follow-up workshop, which will be held in Houston on September 4-5, 2019.
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- 2019
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24. An Interferon-Driven Oxysterol-Based Defense against Tumor-Derived Extracellular Vesicles.
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Ortiz A, Gui J, Zahedi F, Yu P, Cho C, Bhattacharya S, Carbone CJ, Yu Q, Katlinski KV, Katlinskaya YV, Handa S, Haas V, Volk SW, Brice AK, Wals K, Matheson NJ, Antrobus R, Ludwig S, Whiteside TL, Sander C, Tarhini AA, Kirkwood JM, Lehner PJ, Guo W, Rui H, Minn AJ, Koumenis C, Diehl JA, and Fuchs SY
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- Animals, Cell Line, Tumor, Disease Progression, Gene Expression Regulation, Neoplastic drug effects, Gene Knockout Techniques, Humans, Interferons pharmacology, Lung Neoplasms metabolism, Lung Neoplasms pathology, Melanoma metabolism, Mice, Neoplasm Metastasis, Oxysterols metabolism, Reserpine administration & dosage, Reserpine pharmacology, Steroid Hydroxylases genetics, THP-1 Cells, Extracellular Vesicles metabolism, Lung Neoplasms secondary, Melanoma pathology, Receptor, Interferon alpha-beta metabolism, Steroid Hydroxylases metabolism
- Abstract
Tumor-derived extracellular vesicles (TEV) "educate" healthy cells to promote metastases. We found that melanoma TEV downregulated type I interferon (IFN) receptor and expression of IFN-inducible cholesterol 25-hydroxylase (CH25H). CH25H produces 25-hydroxycholesterol, which inhibited TEV uptake. Low CH25H levels in leukocytes from melanoma patients correlated with poor prognosis. Mice incapable of downregulating the IFN receptor and Ch25h were resistant to TEV uptake, TEV-induced pre-metastatic niche, and melanoma lung metastases; however, ablation of Ch25h reversed these phenotypes. An anti-hypertensive drug, reserpine, suppressed TEV uptake and disrupted TEV-induced formation of the pre-metastatic niche and melanoma lung metastases. These results suggest the importance of CH25H in defense against education of normal cells by TEV and argue for the use of reserpine in adjuvant melanoma therapy., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2019
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25. A phase I trial of pembrolizumab with hypofractionated radiotherapy in patients with metastatic solid tumours.
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Maity A, Mick R, Huang AC, George SM, Farwell MD, Lukens JN, Berman AT, Mitchell TC, Bauml J, Schuchter LM, O'Hara M, Lin LL, Demichele A, Christodouleas JP, Haas NB, Patsch DM, Hahn SM, Minn AJ, Wherry EJ, and Vonderheide RH
- Subjects
- Adult, Aged, Aged, 80 and over, Carcinoma, Non-Small-Cell Lung pathology, Disease-Free Survival, Female, Humans, Lung Neoplasms pathology, Male, Melanoma pathology, Middle Aged, Neoplasm Metastasis drug therapy, Neoplasm Metastasis radiotherapy, Skin Neoplasms pathology, Antibodies, Monoclonal, Humanized therapeutic use, Antineoplastic Agents, Immunological therapeutic use, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung radiotherapy, Chemoradiotherapy, Lung Neoplasms drug therapy, Lung Neoplasms radiotherapy, Melanoma drug therapy, Melanoma radiotherapy, Radiation Dose Hypofractionation, Skin Neoplasms drug therapy, Skin Neoplasms radiotherapy
- Abstract
Background: We conducted a phase I trial evaluating pembrolizumab+hypofractionated radiotherapy (HFRT) for patients with metastatic cancers., Methods: There were two strata (12 patients each): (i) NSCLC/melanoma progressing on prior anti-PD-1 therapy, (ii) other cancer types; anti-PD-1-naive. Patients received 6 cycles of pembrolizumab, starting 1 week before HFRT. Patients had ≥2 lesions; only one was irradiated (8 Gy × 3 for first half; 17 Gy × 1 for second half in each stratum) and the other(s) followed for response., Results: Of the 24 patients, 20 (83%) had treatment-related adverse events (AEs) (all grade 1 or 2). There were eight grade 3 AEs, none treatment related. There were no dose-limiting toxicities or grade 4/5 AEs. Stratum 1: two patients (of 12) with progression on prior PD-1 blockade experienced prolonged responses (9.2 and 28.1 months). Stratum 2: one patient experienced a complete response and two had prolonged stable disease (7.4 and 7.0 months). Immune profiling demonstrated that anti-PD-1 therapy and radiation induced a consistent increase in the proliferation marker Ki67 in PD-1-expressing CD8 T cells., Conclusions: HFRT was well tolerated with pembrolizumab, and in some patients with metastatic NSCLC or melanoma, it reinvigorated a systemic response despite previous progression on anti-PD-1 therapy., Clinical Trial Registration: NCT02303990 ( www.clinicaltrials.gov ).
- Published
- 2018
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26. Radiotherapy and CD40 Activation Separately Augment Immunity to Checkpoint Blockade in Cancer.
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Rech AJ, Dada H, Kotzin JJ, Henao-Mejia J, Minn AJ, Twyman-Saint Victor C, and Vonderheide RH
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- Adaptive Immunity immunology, Animals, Antigen-Presenting Cells immunology, CD8-Positive T-Lymphocytes immunology, Carcinoma, Pancreatic Ductal therapy, Cell Line, Tumor, Female, Immune Tolerance immunology, Immunotherapy methods, Immunotherapy, Adoptive methods, Male, Mice, Mice, Inbred C57BL, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms therapy, Tumor Microenvironment immunology, Pancreatic Neoplasms, CD40 Antigens immunology, Carcinoma, Pancreatic Ductal immunology, Carcinoma, Pancreatic Ductal radiotherapy, Pancreatic Neoplasms immunology, Pancreatic Neoplasms radiotherapy
- Abstract
Immunotherapy in pancreatic ductal adenocarcinoma (PDA) remains a difficult clinical problem despite success in other disease types with immune checkpoint blockade (ICB) and chimeric antigen receptor T-cell therapy. Mechanisms driving immunosuppression and poor T-cell infiltration in PDA are incompletely understood. Here, we use genetically engineered mouse models of PDA that recapitulate hallmarks of human disease to demonstrate that CD40 pathway activation is required for clinical response to radiotherapy and ICB with αCTLA-4 and αPD-1. The combination of an agonist αCD40 antibody, radiotherapy, and dual ICB eradicated irradiated and unirradiated (i.e., abscopal) tumors, generating long-term immunity. Response required T cells and also short-lived myeloid cells and was dependent on the long noncoding RNA myeloid regulator Morrbid Using unbiased random forest machine learning, we built unique, contextual signatures for each therapeutic component, revealing that (i) radiotherapy triggers an early proinflammatory stimulus, ablating existing intratumoral T cells and upregulating MHC class I and CD86 on antigen-presenting cells, (ii) αCD40 causes a systemic and intratumoral reorganization of the myeloid compartment, and (iii) ICB increases intratumoral T-cell infiltration and improves the CD8 T-cell:regulatory T-cell ratio. Thus, αCD40 and radiotherapy nonredundantly augment antitumor immunity in PDA, which is otherwise refractory to ICB, providing a clear rationale for clinical evaluation. Significance: Radiotherapy and αCD40 disrupt key links between innate and adaptive immunity, ameliorating resistance to immune checkpoint blockade in pancreatic cancer via multiple cellular mechanisms. Cancer Res; 78(15); 4282-91. ©2018 AACR ., (©2018 American Association for Cancer Research.)
- Published
- 2018
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27. Combination Cancer Therapy with Immune Checkpoint Blockade: Mechanisms and Strategies.
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Patel SA and Minn AJ
- Subjects
- Animals, Antineoplastic Agents, Immunological pharmacology, Antineoplastic Agents, Immunological therapeutic use, Biomarkers, Tumor, Combined Modality Therapy, Humans, Immune System cytology, Immune System drug effects, Immune System immunology, Immune System metabolism, Immunomodulation drug effects, Neoplasms metabolism, Neoplasms pathology, Receptors, Pattern Recognition metabolism, Signal Transduction drug effects, Tumor Microenvironment drug effects, Tumor Microenvironment immunology, Immunotherapy, Molecular Targeted Therapy, Neoplasms immunology, Neoplasms therapy
- Abstract
The success of immune checkpoint blockade in patients with a wide variety of malignancies has changed the treatment paradigm in oncology. However, combination therapies with immune checkpoint blockade will be needed to overcome resistance and broaden the clinical utility of immunotherapy. Here we discuss a framework for rationally designing combination therapy strategies based on enhancing major discriminatory functions of the immune system that are corrupted by cancer-namely, antigenicity, adjuvanticity, and homeostatic feedback inhibition. We review recent advances on how conventional genotoxic cancer therapies, molecularly targeted therapies, epigenetic agents, and immune checkpoint inhibitors can restore these discriminatory functions. Potential barriers that can impede response despite combination therapy are also discussed., (Copyright © 2018. Published by Elsevier Inc.)
- Published
- 2018
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28. Survivorship care planning in skin cancer: An unbiased statistical approach to identifying patterns of care-plan use.
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Benci JL, Minn AJ, Vachani CC, Bach C, Arnold-Korzeniowski K, Hampshire MK, Metz JM, and Hill-Kayser CE
- Subjects
- Academic Medical Centers, Adult, Age Factors, Aged, Aged, 80 and over, Female, Humans, Male, Middle Aged, Oncologists, Physicians, Primary Care, Supervised Machine Learning, United States, Aftercare methods, Cancer Survivors, Melanoma therapy, Patient Care Planning statistics & numerical data, Skin Neoplasms therapy, Survivorship
- Abstract
Background: Nearly 1 in 5 Americans will develop skin cancer, and as a result, survivors of skin cancer compose one of the largest groups of cancer survivors. Survivorship care plans (SCPs) are an important tool for improving patient outcomes and provide critical information to both survivors and health care professionals. Recent efforts have been made to expand SCP utilization; however, which patients currently receive SCPs is poorly understood., Methods: This study used 596 individuals with a diagnosis of melanoma (n = 391) or nonmelanoma skin cancer (n = 205) who had used an Internet-based SCP tool from May 2010 to December 2016 to model the patient and provider characteristics that determine SCP utilization., Results: Survivors were predominantly white (95.3%) and female (56.5%). Survivors who received a treatment summary were more likely to also receive an SCP. University and nonuniversity cancer centers used SCPs at a higher rate than other care settings. Survivors whose care was managed by a team rather than just an individual physician were also more likely to receive an SCP. Survivors older than 70 years at diagnosis were almost twice as likely to receive a plan as survivors who were diagnosed at a younger age., Conclusions: With a convenience sample of skin cancer survivors, it is possible to model factors that predict the receipt of SCPs. Important variables include the diagnosis age, treatment setting, physician type, and treatment-summary utilization. A closer examination of these variables identified several disparities in care-plan use and, therefore, opportunities to improve the distribution of SCPs. Further validation in additional cohorts of survivors is necessary to confirm these conclusions. Cancer 2018;124:183-91. © 2017 American Cancer Society., (© 2017 American Cancer Society.)
- Published
- 2018
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29. Mitotic progression following DNA damage enables pattern recognition within micronuclei.
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Harding SM, Benci JL, Irianto J, Discher DE, Minn AJ, and Greenberg RA
- Subjects
- Animals, CTLA-4 Antigen antagonists & inhibitors, Cell Cycle Checkpoints, Cell Line, Tumor, DNA Breaks, Double-Stranded, Disease Models, Animal, Female, Humans, Inflammation pathology, Interferons metabolism, Melanoma, Experimental drug therapy, Melanoma, Experimental metabolism, Melanoma, Experimental pathology, Membrane Proteins metabolism, Mice, Mice, Inbred C57BL, Nucleotidyltransferases metabolism, DNA Damage, Inflammation metabolism, Micronuclei, Chromosome-Defective, Mitosis, Receptors, Pattern Recognition metabolism, Signal Transduction
- Abstract
Inflammatory gene expression following genotoxic cancer therapy is well documented, yet the events underlying its induction remain poorly understood. Inflammatory cytokines modify the tumour microenvironment by recruiting immune cells and are critical for both local and systemic (abscopal) tumour responses to radiotherapy. A poorly understood feature of these responses is the delayed onset (days), in contrast to the acute DNA-damage responses that occur in minutes to hours. Such dichotomous kinetics implicate additional rate-limiting steps that are essential for DNA-damage-induced inflammation. Here we show that cell cycle progression through mitosis following double-stranded DNA breaks leads to the formation of micronuclei, which precede activation of inflammatory signalling and are a repository for the pattern-recognition receptor cyclic GMP-AMP synthase (cGAS). Inhibiting progression through mitosis or loss of pattern recognition by stimulator of interferon genes (STING)-cGAS impaired interferon signalling. Moreover, STING loss prevented the regression of abscopal tumours in the context of ionizing radiation and immune checkpoint blockade in vivo. These findings implicate temporal modulation of the cell cycle as an important consideration in the context of therapeutic strategies that combine genotoxic agents with immune checkpoint blockade.
- Published
- 2017
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30. Nuclear Acetyl-CoA Production by ACLY Promotes Homologous Recombination.
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Sivanand S, Rhoades S, Jiang Q, Lee JV, Benci J, Zhang J, Yuan S, Viney I, Zhao S, Carrer A, Bennett MJ, Minn AJ, Weljie AM, Greenberg RA, and Wellen KE
- Subjects
- A549 Cells, ATP Citrate (pro-S)-Lyase genetics, Acetylation, Animals, BRCA1 Protein genetics, Cell Nucleus drug effects, Female, G2 Phase Cell Cycle Checkpoints, Genomic Instability, Glucose metabolism, HCT116 Cells, HeLa Cells, Histones metabolism, Humans, Melanoma, Experimental enzymology, Melanoma, Experimental genetics, Melanoma, Experimental pathology, Mice, Inbred C57BL, Phosphorylation, Poly(ADP-ribose) Polymerase Inhibitors pharmacology, Protein Binding, Protein Processing, Post-Translational, RNA Interference, S Phase Cell Cycle Checkpoints, Serine, Time Factors, Transfection, Tumor Suppressor p53-Binding Protein 1 metabolism, ATP Citrate (pro-S)-Lyase metabolism, Acetyl Coenzyme A metabolism, BRCA1 Protein metabolism, Cell Nucleus enzymology, DNA Breaks, Double-Stranded, Recombinational DNA Repair drug effects
- Abstract
While maintaining the integrity of the genome and sustaining bioenergetics are both fundamental functions of the cell, potential crosstalk between metabolic and DNA repair pathways is poorly understood. Since histone acetylation plays important roles in DNA repair and is sensitive to the availability of acetyl coenzyme A (acetyl-CoA), we investigated a role for metabolic regulation of histone acetylation during the DNA damage response. In this study, we report that nuclear ATP-citrate lyase (ACLY) is phosphorylated at S455 downstream of ataxia telangiectasia mutated (ATM) and AKT following DNA damage. ACLY facilitates histone acetylation at double-strand break (DSB) sites, impairing 53BP1 localization and enabling BRCA1 recruitment and DNA repair by homologous recombination. ACLY phosphorylation and nuclear localization are necessary for its role in promoting BRCA1 recruitment. Upon PARP inhibition, ACLY silencing promotes genomic instability and cell death. Thus, the spatial and temporal control of acetyl-CoA production by ACLY participates in the mechanism of DNA repair pathway choice., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
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31. Exosome RNA Unshielding Couples Stromal Activation to Pattern Recognition Receptor Signaling in Cancer.
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Nabet BY, Qiu Y, Shabason JE, Wu TJ, Yoon T, Kim BC, Benci JL, DeMichele AM, Tchou J, Marcotrigiano J, and Minn AJ
- Subjects
- Breast Neoplasms metabolism, DEAD Box Protein 58 metabolism, Exosomes metabolism, Humans, Interferon Regulatory Factors metabolism, MCF-7 Cells, Neoplasm Metastasis, RNA Polymerase III genetics, RNA Polymerase III metabolism, Receptors, Immunologic, Receptors, Pattern Recognition metabolism, Signal Recognition Particle metabolism, Stromal Cells metabolism, Virus Diseases metabolism, Breast Neoplasms pathology, Exosomes pathology, RNA, Untranslated metabolism, Stromal Cells pathology, Tumor Microenvironment
- Abstract
Interactions between stromal fibroblasts and cancer cells generate signals for cancer progression, therapy resistance, and inflammatory responses. Although endogenous RNAs acting as damage-associated molecular patterns (DAMPs) for pattern recognition receptors (PRRs) may represent one such signal, these RNAs must remain unrecognized under non-pathological conditions. We show that triggering of stromal NOTCH-MYC by breast cancer cells results in a POL3-driven increase in RN7SL1, an endogenous RNA normally shielded by RNA binding proteins SRP9/14. This increase in RN7SL1 alters its stoichiometry with SRP9/14 and generates unshielded RN7SL1 in stromal exosomes. After exosome transfer to immune cells, unshielded RN7SL1 drives an inflammatory response. Upon transfer to breast cancer cells, unshielded RN7SL1 activates the PRR RIG-I to enhance tumor growth, metastasis, and therapy resistance. Corroborated by evidence from patient tumors and blood, these results demonstrate that regulation of RNA unshielding couples stromal activation with deployment of RNA DAMPs that promote aggressive features of cancer. VIDEO ABSTRACT., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
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32. Radiation and Immune Checkpoint Blockade: From Bench to Clinic.
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Shabason JE and Minn AJ
- Subjects
- Combined Modality Therapy methods, Humans, Neoplasm Metastasis, Tumor Escape, Immunotherapy methods, Neoplasms immunology, Neoplasms therapy
- Abstract
Immune escape of malignant cells is an important hallmark of cancer, necessary for tumor formation and progression. Accordingly, in recent years, therapies that enhance the immune system have had remarkable success in treating a myriad of malignancies. Particularly successful has been immune checkpoint blockade (ICB), which is a therapy that targets T-cell inhibitory receptors, or immune checkpoints. Despite these encouraging clinical results, most patients do not respond to such agents. Therefore, determining methods to better target and enhance the therapeutic efficacy of ICB is of paramount importance. One appealing approach is to use standard anticancer therapies, such as radiation, chemotherapy, and targeted biologics, to favorably modulate the immune system and enhance the anticancer immune response. For example, although radiation therapy has classically been thought of as a local therapy, there is significant potential for combining radiation therapy with ICB to both optimize local control and to treat metastatic disease. This concept is supported by numerous preclinical studies and clinical case reports and has since led to many early and ongoing clinical trials. However, it is still unclear how to optimally combine radiation and ICB to maximize the therapeutic effect. In this review, we highlight relevant preclinical and clinical studies in the field of radiation and ICB and discuss optimal strategies for combination therapies moving forward., (Copyright © 2017. Published by Elsevier Inc.)
- Published
- 2017
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33. Tumor Interferon Signaling Regulates a Multigenic Resistance Program to Immune Checkpoint Blockade.
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Benci JL, Xu B, Qiu Y, Wu TJ, Dada H, Twyman-Saint Victor C, Cucolo L, Lee DSM, Pauken KE, Huang AC, Gangadhar TC, Amaravadi RK, Schuchter LM, Feldman MD, Ishwaran H, Vonderheide RH, Maity A, Wherry EJ, and Minn AJ
- Subjects
- Animals, B7-H1 Antigen metabolism, Cell Line, Tumor, Drug Resistance, Neoplasm, Heterografts, Humans, Interferons immunology, Melanoma drug therapy, Melanoma radiotherapy, Mice, Neoplasm Transplantation, STAT1 Transcription Factor, T-Lymphocytes immunology, CTLA-4 Antigen antagonists & inhibitors, Melanoma immunology, Melanoma therapy, Radioimmunotherapy
- Abstract
Therapeutic blocking of the PD1 pathway results in significant tumor responses, but resistance is common. We demonstrate that prolonged interferon signaling orchestrates PDL1-dependent and PDL1-independent resistance to immune checkpoint blockade (ICB) and to combinations such as radiation plus anti-CTLA4. Persistent type II interferon signaling allows tumors to acquire STAT1-related epigenomic changes and augments expression of interferon-stimulated genes and ligands for multiple T cell inhibitory receptors. Both type I and II interferons maintain this resistance program. Crippling the program genetically or pharmacologically interferes with multiple inhibitory pathways and expands distinct T cell populations with improved function despite expressing markers of severe exhaustion. Consequently, tumors resistant to multi-agent ICB are rendered responsive to ICB monotherapy. Finally, we observe that biomarkers for interferon-driven resistance associate with clinical progression after anti-PD1 therapy. Thus, the duration of tumor interferon signaling augments adaptive resistance and inhibition of the interferon response bypasses requirements for combinatorial ICB therapies., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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34. Assessing intratumor heterogeneity and tracking longitudinal and spatial clonal evolutionary history by next-generation sequencing.
- Author
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Jiang Y, Qiu Y, Minn AJ, and Zhang NR
- Subjects
- Computational Biology methods, DNA Copy Number Variations genetics, Exome genetics, Genetic Heterogeneity, Humans, Internet, Mutation, Neoplasms pathology, Phylogeny, Polymorphism, Single Nucleotide, Software, Clonal Evolution genetics, Evolution, Molecular, High-Throughput Nucleotide Sequencing methods, Neoplasms genetics
- Abstract
Cancer is a disease driven by evolutionary selection on somatic genetic and epigenetic alterations. Here, we propose Canopy, a method for inferring the evolutionary phylogeny of a tumor using both somatic copy number alterations and single-nucleotide alterations from one or more samples derived from a single patient. Canopy is applied to bulk sequencing datasets of both longitudinal and spatial experimental designs and to a transplantable metastasis model derived from human cancer cell line MDA-MB-231. Canopy successfully identifies cell populations and infers phylogenies that are in concordance with existing knowledge and ground truth. Through simulations, we explore the effects of key parameters on deconvolution accuracy and compare against existing methods. Canopy is an open-source R package available at https://cran.r-project.org/web/packages/Canopy/., Competing Interests: The authors declare no conflict of interest.
- Published
- 2016
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35. Combination Cancer Therapies with Immune Checkpoint Blockade: Convergence on Interferon Signaling.
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Minn AJ and Wherry EJ
- Subjects
- Animals, Humans, Immune Tolerance, Immunity, Innate, Lymphocytes immunology, Neoplasms metabolism, Interferons metabolism, Neoplasms immunology, Neoplasms therapy, Signal Transduction
- Abstract
Improving efficacy of immune checkpoint blockade for cancer can be facilitated by combining these agents with each other and/or with other conventional or targeted therapies. Interferon and innate immune signaling pathways in immune and tumor cells have emerged as intriguing determinants of response and resistance, often in complex and seemingly paradoxical ways., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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36. Erratum to: PI3K/mTOR inhibition can impair tumor invasion and metastasis in vivo despite a lack of antiproliferative action in vitro: implications for targeted therapy.
- Author
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Wander SA, Zhao D, Besser AH, Hong F, Wei J, Ince TA, Milikowski C, Bishopric NH, Minn AJ, Creighton CJ, and Slingerland JM
- Abstract
Erratum to: Breast Cancer Res Treat (2013),138:369–381,DOI 10.1007/s10549-012-2389-6. In the original publication of the article, the Fig. 4c and d were published erroneously. The revised Fig. 4 is given in this erratum.
- Published
- 2016
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37. Getting Tumor Dendritic Cells to Engage the Dead.
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Cucolo L and Minn AJ
- Subjects
- Animals, Female, Humans, Anthracyclines therapeutic use, Neoplasms drug therapy, Neoplasms immunology, Receptors, Formyl Peptide physiology
- Abstract
The immunogenic effects of chemotherapy rely on effective activation of dendritic cells to present antigen to tumor-specific T cells. However, the signals that govern how dendritic cells seek out dying cancer cells to initiate this process are poorly understood. A recent study by Vacchelli et al. provides important insight., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
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38. Awakening the immune system with radiation: Optimal dose and fractionation.
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Gandhi SJ, Minn AJ, Vonderheide RH, Wherry EJ, Hahn SM, and Maity A
- Subjects
- Animals, Dose Fractionation, Radiation, Dose-Response Relationship, Radiation, Humans, Radiotherapy methods, Immune System radiation effects, Neoplasms immunology, Neoplasms radiotherapy
- Abstract
The importance of ionizing radiation has historically been limited to achieving local control of tumor cells. However, emerging evidence over the last decade suggests an increasingly important role for radiation in amplifying the antitumor immune response elicited by immunomodulatory agents. Combination of radiation with immunotherapy has been shown to elicit powerful systemic responses in several pre-clinical tumor models. Additionally, recent clinical observations support the use of radiation therapy for augmenting antitumor immunity in the metastatic setting. However, radiation dose and fractionation schedules for optimal synergy between radiotherapy and immunotherapy are not well defined. Here we review pre-clinical and clinical data relating to radiation dose and fractionation in the setting of immunotherapy and discuss optimal strategies for combining the two therapies., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2015
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39. Tumour exosome integrins determine organotropic metastasis.
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Hoshino A, Costa-Silva B, Shen TL, Rodrigues G, Hashimoto A, Tesic Mark M, Molina H, Kohsaka S, Di Giannatale A, Ceder S, Singh S, Williams C, Soplop N, Uryu K, Pharmer L, King T, Bojmar L, Davies AE, Ararso Y, Zhang T, Zhang H, Hernandez J, Weiss JM, Dumont-Cole VD, Kramer K, Wexler LH, Narendran A, Schwartz GK, Healey JH, Sandstrom P, Labori KJ, Kure EH, Grandgenett PM, Hollingsworth MA, de Sousa M, Kaur S, Jain M, Mallya K, Batra SK, Jarnagin WR, Brady MS, Fodstad O, Muller V, Pantel K, Minn AJ, Bissell MJ, Garcia BA, Kang Y, Rajasekhar VK, Ghajar CM, Matei I, Peinado H, Bromberg J, and Lyden D
- Subjects
- Animals, Biomarkers metabolism, Brain cytology, Cell Line, Tumor, Endothelial Cells cytology, Endothelial Cells metabolism, Epithelial Cells cytology, Epithelial Cells metabolism, Female, Fibroblasts cytology, Fibroblasts metabolism, Genes, src, Humans, Integrin alpha6beta1 metabolism, Integrin alpha6beta4 antagonists & inhibitors, Integrin alpha6beta4 metabolism, Integrin beta Chains metabolism, Integrin beta4 metabolism, Integrins antagonists & inhibitors, Kupffer Cells cytology, Kupffer Cells metabolism, Liver cytology, Lung cytology, Mice, Mice, Inbred C57BL, Organ Specificity, Phosphorylation, Receptors, Vitronectin antagonists & inhibitors, Receptors, Vitronectin metabolism, S100 Proteins genetics, Brain metabolism, Exosomes metabolism, Integrins metabolism, Liver metabolism, Lung metabolism, Neoplasm Metastasis pathology, Neoplasm Metastasis prevention & control, Tropism
- Abstract
Ever since Stephen Paget's 1889 hypothesis, metastatic organotropism has remained one of cancer's greatest mysteries. Here we demonstrate that exosomes from mouse and human lung-, liver- and brain-tropic tumour cells fuse preferentially with resident cells at their predicted destination, namely lung fibroblasts and epithelial cells, liver Kupffer cells and brain endothelial cells. We show that tumour-derived exosomes uptaken by organ-specific cells prepare the pre-metastatic niche. Treatment with exosomes from lung-tropic models redirected the metastasis of bone-tropic tumour cells. Exosome proteomics revealed distinct integrin expression patterns, in which the exosomal integrins α6β4 and α6β1 were associated with lung metastasis, while exosomal integrin αvβ5 was linked to liver metastasis. Targeting the integrins α6β4 and αvβ5 decreased exosome uptake, as well as lung and liver metastasis, respectively. We demonstrate that exosome integrin uptake by resident cells activates Src phosphorylation and pro-inflammatory S100 gene expression. Finally, our clinical data indicate that exosomal integrins could be used to predict organ-specific metastasis.
- Published
- 2015
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40. Interferons and the Immunogenic Effects of Cancer Therapy.
- Author
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Minn AJ
- Subjects
- Animals, Humans, Neoplasms pathology, Interferons immunology, Neoplasms drug therapy, Neoplasms immunology
- Abstract
Much of our understanding on resistance mechanisms to conventional cancer therapies such as chemotherapy and radiation has focused on cell intrinsic properties that antagonize the detrimental effects of DNA and other cellular damage. However, it is becoming clear that the immune system and/or innate immune signaling pathways can integrate with these intrinsic mechanisms to profoundly influence treatment efficacy. In this context, recent evidence indicates that interferon (IFN) signaling has an important role in this integration by influencing immune and intrinsic/non-immune determinants of therapy response. However, IFN signaling can be both immunostimulatory and immunosuppressive, and the factors determining these outcomes in different disease settings are unclear. Here I discuss the regulation and molecular events in cancer that are associated with these dichotomous functions.
- Published
- 2015
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41. αB-crystallin Expression in Breast Cancer is Associated with Brain Metastasis.
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Voduc KD, Nielsen TO, Perou CM, Harrell JC, Fan C, Kennecke H, Minn AJ, Cryns VL, and Cheang MCU
- Abstract
Background/objectives: The molecular chaperone αB-crystallin is expressed in estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2 "triple-negative" breast carcinomas and promotes brain and lung metastasis. We examined αB-crystallin expression in primary breast carcinomas with metastatic data to evaluate its association with prognosis and site-specific metastases., Methods: αB-crystallin gene ( CRYAB ) expression was examined using publically available global-gene expression data (n=855 breast tumors) with first site of distant metastasis information ("855Met"). αB-crystallin protein expression was determined by immunohistochemistry using the clinically annotated tissue microarray (n=3987 breast tumors) from British Columbia Cancer Agency (BCCA). Kaplan-Meier and multivariable Cox regression analyses were used to evaluate the prognostic value of αB-crystallin. Multivariable logistic regression analysis was used to evaluate risks of αB-crystallin and other markers for site of metastasis., Results: In the 855Met dataset, αB-crystallin gene ( CRYAB) expression was an independent predictor of brain as the first distant site of relapse (HR = 1.2, (95% CI 1.0-1.4), P = 0.021). In the BCCA series, αB-crystallin protein expression was an independent prognostic marker of poor breast cancer specific survival (HR = 1.3, (95% CI 1.1-1.6), P = 0.014). Among patients with metastases, αB-crystallin was the strongest independent predictor of brain metastasis (OR = 2.99 (95% CI 1.83-4.89), P < 0.0001) and the only independent predictor of brain as the first site of distant metastasis (OR = 3.15 (95% CI1.43-6.95), P = 0.005). αB-crystallin was also associated with worse survival (3.0 versus 4.7 months, P = 0.007)., Conclusions: αB-crystallin is a promising biomarker to identify breast cancer patients at high risk for early relapse in the brain, independent of ER and HER2 status., Competing Interests: Competing interests CMP and TON are equity stock holders, and Board of Director Member of BioClassifier LLC. CMP, TON and MCUC are also listed as inventors on a patent application on the PAM50 assay. The other authors declare they have no competing interests.
- Published
- 2015
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42. Linking primary and metastatic tumour re-initiation.
- Author
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Nabet BY and Minn AJ
- Subjects
- Animals, Female, Humans, Biomarkers, Tumor metabolism, Breast Neoplasms metabolism, Cell Proliferation, Laminin metabolism, Liver Neoplasms metabolism, Lung Neoplasms metabolism, Neoplastic Stem Cells metabolism
- Abstract
Whether the cancer cells responsible for the growth of primary tumours are also able to re-initiate tumour growth after seeding to distant organs is unclear. The characterization of breast cancer cells with both of these attributes now identifies the functional and molecular determinants necessary to mediate primary tumour formation and re-initiation at the secondary site.
- Published
- 2015
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43. MicroRNA-124 expression counteracts pro-survival stress responses in glioblastoma.
- Author
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Mucaj V, Lee SS, Skuli N, Giannoukos DN, Qiu B, Eisinger-Mathason TS, Nakazawa MS, Shay JE, Gopal PP, Venneti S, Lal P, Minn AJ, Simon MC, and Mathew LK
- Subjects
- Animals, Brain Neoplasms genetics, Brain Neoplasms pathology, Cell Hypoxia, Cell Line, Tumor, Cell Survival, Glioblastoma genetics, Glioblastoma pathology, Heterografts, Humans, Mice, Mice, Nude, MicroRNAs genetics, Neoplasm Proteins, Neoplasm Transplantation, RNA, Neoplasm genetics, Brain Neoplasms metabolism, Gene Expression Regulation, Neoplastic, Glioblastoma metabolism, MicroRNAs biosynthesis, RNA, Neoplasm biosynthesis, Stress, Physiological
- Abstract
Glioblastomas are aggressive adult brain tumors, characterized by inadequately organized vasculature and consequent nutrient and oxygen (O2)-depleted areas. Adaptation to low nutrients and hypoxia supports glioblastoma cell survival, progression and therapeutic resistance. However, specific mechanisms promoting cellular survival under nutrient and O2 deprivation remain incompletely understood. Here, we show that miR-124 expression is negatively correlated with a hypoxic gene signature in glioblastoma patient samples, suggesting that low miR-124 levels contribute to pro-survival adaptive pathways in this disease. As miR-124 expression is repressed in various cancer types (including glioblastoma), we quantified miR-124 abundance in normoxic and hypoxic regions in glioblastoma patient tissue, and investigated whether ectopic miR-124 expression compromises cell survival during tumor ischemia. Our results indicate that miR-124 levels are further diminished in hypoxic/ischemic regions within individual glioblastoma patient samples, compared with regions replete in O2 and nutrients. Importantly, we also show that increased miR-124 expression affects the ability of tumor cells to survive under O2 and/or nutrient deprivation. Moreover, miR-124 re-expression increases cell death in vivo and enhances the survival of mice bearing intracranial xenograft tumors. miR-124 exerts this phenotype in part by directly regulating TEAD1, MAPK14/p38α and SERP1, factors involved in cell proliferation and survival under stress. Simultaneous suppression of these miR-124 targets results in similar levels of cell death as caused by miR-124 restoration. Importantly, we further demonstrate that SERP1 reintroduction reverses the hypoxic cell death elicited by miR-124, indicating the importance of SERP1 in promoting tumor cell survival. In support of our experimental data, we observed a significant correlation between high SERP1 levels and poor patient outcome in glioblastoma patients. Collectively, among the many pro-tumorigeneic properties of miR-124 repression in glioblastoma, we delineated a novel role in promoting tumor cell survival under stressful microenvironments, thereby supporting tumor progression.
- Published
- 2015
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44. Radiation and dual checkpoint blockade activate non-redundant immune mechanisms in cancer.
- Author
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Twyman-Saint Victor C, Rech AJ, Maity A, Rengan R, Pauken KE, Stelekati E, Benci JL, Xu B, Dada H, Odorizzi PM, Herati RS, Mansfield KD, Patsch D, Amaravadi RK, Schuchter LM, Ishwaran H, Mick R, Pryma DA, Xu X, Feldman MD, Gangadhar TC, Hahn SM, Wherry EJ, Vonderheide RH, and Minn AJ
- Subjects
- Animals, B7-H1 Antigen metabolism, Female, Humans, Melanoma pathology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Receptors, Antigen, T-Cell drug effects, Receptors, Antigen, T-Cell immunology, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes cytology, T-Lymphocytes immunology, T-Lymphocytes, Regulatory drug effects, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory radiation effects, B7-H1 Antigen antagonists & inhibitors, CTLA-4 Antigen antagonists & inhibitors, Cell Cycle Checkpoints drug effects, Melanoma drug therapy, Melanoma immunology, Melanoma radiotherapy, T-Lymphocytes drug effects, T-Lymphocytes radiation effects
- Abstract
Immune checkpoint inhibitors result in impressive clinical responses, but optimal results will require combination with each other and other therapies. This raises fundamental questions about mechanisms of non-redundancy and resistance. Here we report major tumour regressions in a subset of patients with metastatic melanoma treated with an anti-CTLA4 antibody (anti-CTLA4) and radiation, and reproduced this effect in mouse models. Although combined treatment improved responses in irradiated and unirradiated tumours, resistance was common. Unbiased analyses of mice revealed that resistance was due to upregulation of PD-L1 on melanoma cells and associated with T-cell exhaustion. Accordingly, optimal response in melanoma and other cancer types requires radiation, anti-CTLA4 and anti-PD-L1/PD-1. Anti-CTLA4 predominantly inhibits T-regulatory cells (Treg cells), thereby increasing the CD8 T-cell to Treg (CD8/Treg) ratio. Radiation enhances the diversity of the T-cell receptor (TCR) repertoire of intratumoral T cells. Together, anti-CTLA4 promotes expansion of T cells, while radiation shapes the TCR repertoire of the expanded peripheral clones. Addition of PD-L1 blockade reverses T-cell exhaustion to mitigate depression in the CD8/Treg ratio and further encourages oligoclonal T-cell expansion. Similarly to results from mice, patients on our clinical trial with melanoma showing high PD-L1 did not respond to radiation plus anti-CTLA4, demonstrated persistent T-cell exhaustion, and rapidly progressed. Thus, PD-L1 on melanoma cells allows tumours to escape anti-CTLA4-based therapy, and the combination of radiation, anti-CTLA4 and anti-PD-L1 promotes response and immunity through distinct mechanisms.
- Published
- 2015
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45. Exosome transfer from stromal to breast cancer cells regulates therapy resistance pathways.
- Author
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Boelens MC, Wu TJ, Nabet BY, Xu B, Qiu Y, Yoon T, Azzam DJ, Twyman-Saint Victor C, Wiemann BZ, Ishwaran H, Ter Brugge PJ, Jonkers J, Slingerland J, and Minn AJ
- Subjects
- Animals, Breast Neoplasms metabolism, Cell Line, Tumor, Computer Simulation, Drug Resistance, Neoplasm, Female, Humans, Interferons metabolism, Mice, Nude, Radiation Tolerance, Receptors, Notch metabolism, STAT1 Transcription Factor metabolism, Signal Transduction, rab GTP-Binding Proteins metabolism, Breast Neoplasms drug therapy, Breast Neoplasms radiotherapy, Exosomes metabolism, Paracrine Communication, Stromal Cells metabolism
- Abstract
Stromal communication with cancer cells can influence treatment response. We show that stromal and breast cancer (BrCa) cells utilize paracrine and juxtacrine signaling to drive chemotherapy and radiation resistance. Upon heterotypic interaction, exosomes are transferred from stromal to BrCa cells. RNA within exosomes, which are largely noncoding transcripts and transposable elements, stimulates the pattern recognition receptor RIG-I to activate STAT1-dependent antiviral signaling. In parallel, stromal cells also activate NOTCH3 on BrCa cells. The paracrine antiviral and juxtacrine NOTCH3 pathways converge as STAT1 facilitates transcriptional responses to NOTCH3 and expands therapy-resistant tumor-initiating cells. Primary human and/or mouse BrCa analysis support the role of antiviral/NOTCH3 pathways in NOTCH signaling and stroma-mediated resistance, which is abrogated by combination therapy with gamma secretase inhibitors. Thus, stromal cells orchestrate an intricate crosstalk with BrCa cells by utilizing exosomes to instigate antiviral signaling. This expands BrCa subpopulations adept at resisting therapy and reinitiating tumor growth.
- Published
- 2014
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46. miR-218 opposes a critical RTK-HIF pathway in mesenchymal glioblastoma.
- Author
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Mathew LK, Skuli N, Mucaj V, Lee SS, Zinn PO, Sathyan P, Imtiyaz HZ, Zhang Z, Davuluri RV, Rao S, Venneti S, Lal P, Lathia JD, Rich JN, Keith B, Minn AJ, and Simon MC
- Subjects
- Adult, Aged, Aged, 80 and over, Animals, Antineoplastic Agents pharmacology, Cell Survival, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Humans, Hypoxia, Mice, Mice, Nude, Middle Aged, Necrosis, Neoplasm Transplantation, Neovascularization, Pathologic, Oligonucleotide Array Sequence Analysis, Proto-Oncogene Mas, Signal Transduction, Young Adult, Basic Helix-Loop-Helix Transcription Factors metabolism, Brain Neoplasms metabolism, Glioblastoma metabolism, Mesoderm metabolism, MicroRNAs metabolism, Receptor Protein-Tyrosine Kinases metabolism
- Abstract
Glioblastoma multiforme (GBM) and the mesenchymal GBM subtype in particular are highly malignant tumors that frequently exhibit regions of severe hypoxia and necrosis. Because these features correlate with poor prognosis, we investigated microRNAs whose expression might regulate hypoxic GBM cell survival and growth. We determined that the expression of microRNA-218 (miR-218) is decreased significantly in highly necrotic mesenchymal GBM, and orthotopic tumor studies revealed that reduced miR-218 levels confer GBM resistance to chemotherapy. Importantly, miR-218 targets multiple components of receptor tyrosine kinase (RTK) signaling pathways, and miR-218 repression increases the abundance and activity of multiple RTK effectors. This elevated RTK signaling also promotes the activation of hypoxia-inducible factor (HIF), most notably HIF2α. We further show that RTK-mediated HIF2α regulation is JNK dependent, via jun proto-oncogene. Collectively, our results identify an miR-218-RTK-HIF2α signaling axis that promotes GBM cell survival and tumor angiogenesis, particularly in necrotic mesenchymal tumors.
- Published
- 2014
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47. The Myc-miR-17-92 axis amplifies B-cell receptor signaling via inhibition of ITIM proteins: a novel lymphomagenic feed-forward loop.
- Author
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Psathas JN, Doonan PJ, Raman P, Freedman BD, Minn AJ, and Thomas-Tikhonenko A
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Calcium metabolism, Cell Line, Humans, Intracellular Signaling Peptides and Proteins genetics, Intracellular Signaling Peptides and Proteins metabolism, Lymphoma, Large B-Cell, Diffuse genetics, Lymphoma, Large B-Cell, Diffuse pathology, MicroRNAs genetics, Protein-Tyrosine Kinases genetics, Protein-Tyrosine Kinases metabolism, Proto-Oncogene Proteins c-myc genetics, RNA, Long Noncoding, Receptors, Fc genetics, Receptors, IgG genetics, Sialic Acid Binding Ig-like Lectin 2 genetics, Signal Transduction physiology, Syk Kinase, Adaptor Proteins, Signal Transducing metabolism, Lymphoma, Large B-Cell, Diffuse metabolism, Membrane Proteins metabolism, MicroRNAs metabolism, Proto-Oncogene Proteins c-myc metabolism, Receptors, Fc metabolism, Receptors, IgG metabolism, Sialic Acid Binding Ig-like Lectin 2 metabolism
- Abstract
The c-Myc oncoprotein regulates >15% of the human transcriptome and a limited number of microRNAs (miRNAs). Here, we establish that in a human B-lymphoid cell line, Myc-repressed, but not Myc-stimulated, genes are significantly enriched for predicted binding sites of Myc-regulated miRNAs, primarily those comprising the Myc-activated miR-17~92 cluster. Notably, gene set enrichment analysis demonstrates that miR-17∼92 is a major regulator of B-cell receptor (BCR) pathway components. Many of them are immunoreceptor tyrosine inhibitory motif (ITIM)-containing proteins, and ITIM proteins CD22 and FCGR2B were found to be direct targets of miR-17∼92. Consistent with the propensity of ITIM proteins to recruit phosphatases, either MYC or miR-17~92 expression was necessary to sustain phosphorylation of spleen tyrosine kinase (SYK) and the B-cell linker protein (BLNK) upon ligation of the BCR. Further downstream, stimulation of the BCR response by miR-17-92 resulted in the enhanced calcium flux and elevated levels of Myc itself. Notably, inhibition of the miR-17~92 cluster in diffuse large B-cell lymphoma (DLBCL) cell lines diminished the BCR response as measured by SYK and BLNK phosphorylation. Conversely, human DLBCLs of the BCR subtype express higher Myc and mir17hg transcript levels than other subtypes. Hence, the Myc-miR-17-92-BCR axis, frequently affected by genomic rearrangements, constitutes a novel lymphomagenic feed-forward loop.
- Published
- 2013
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48. Triple negative breast cancer initiating cell subsets differ in functional and molecular characteristics and in γ-secretase inhibitor drug responses.
- Author
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Azzam DJ, Zhao D, Sun J, Minn AJ, Ranganathan P, Drews-Elger K, Han X, Picon-Ruiz M, Gilbert CA, Wander SA, Capobianco AJ, El-Ashry D, and Slingerland JM
- Subjects
- Aldehyde Dehydrogenase 1 Family, Amyloid Precursor Protein Secretases metabolism, Animals, Antineoplastic Agents chemistry, Antineoplastic Agents therapeutic use, Antineoplastic Agents toxicity, CD24 Antigen metabolism, Cell Proliferation drug effects, Enzyme Inhibitors chemistry, Enzyme Inhibitors therapeutic use, Enzyme Inhibitors toxicity, Female, Humans, Hyaluronan Receptors metabolism, Isoenzymes metabolism, MCF-7 Cells, Mice, Mice, Inbred BALB C, Mice, Nude, Neoplastic Stem Cells cytology, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells metabolism, Receptors, Notch metabolism, Retinal Dehydrogenase metabolism, SOXB1 Transcription Factors antagonists & inhibitors, SOXB1 Transcription Factors genetics, SOXB1 Transcription Factors metabolism, Triple Negative Breast Neoplasms drug therapy, Triple Negative Breast Neoplasms metabolism, Triple Negative Breast Neoplasms pathology, Tumor Cells, Cultured, Amyloid Precursor Protein Secretases antagonists & inhibitors
- Abstract
Increasing evidence suggests that stem-like cells mediate cancer therapy resistance and metastasis. Breast tumour-initiating stem cells (T-ISC) are known to be enriched in CD44(+) CD24(neg/low) cells. Here, we identify two T-ISC subsets within this population in triple negative breast cancer (TNBC) lines and dissociated primary breast cancer cultures: CD44(+) CD24(low+) subpopulation generates CD44(+) CD24(neg) progeny with reduced sphere formation and tumourigenicity. CD44(+) CD24(low+) populations contain subsets of ALDH1(+) and ESA(+) cells, yield more frequent spheres and/or T-ISC in limiting dilution assays, preferentially express metastatic gene signatures and show greater motility, invasion and, in the MDA-MB-231 model, metastatic potential. CD44(+) CD24(low+) but not CD44(+) CD24(neg) express activated Notch1 intracellular domain (N1-ICD) and Notch target genes. We show N1-ICD transactivates SOX2 to increase sphere formation, ALDH1+ and CD44(+) CD24(low+) cells. Gamma secretase inhibitors (GSI) reduced sphere formation and xenograft growth from CD44(+) CD24(low+) cells, but CD44(+) CD24(neg) were resistant. While GSI hold promise for targeting T-ISC, stem cell heterogeneity as observed herein, could limit GSI efficacy. These data suggest a breast T-ISC hierarchy in which distinct pathways drive developmentally related subpopulations with different anti-cancer drug responsiveness., (© 2013 The Authors. Published by John Wiley and Sons, Ltd on behalf of EMBO.)
- Published
- 2013
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49. TAp73 enhances the pentose phosphate pathway and supports cell proliferation.
- Author
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Du W, Jiang P, Mancuso A, Stonestrom A, Brewer MD, Minn AJ, Mak TW, Wu M, and Yang X
- Subjects
- Animals, Blotting, Western, Breast Neoplasms physiopathology, Cell Proliferation, Cells, Cultured, DNA biosynthesis, Disease Models, Animal, Female, Gene Expression Regulation, Neoplastic, Gene Knockdown Techniques, Glucosephosphate Dehydrogenase metabolism, Humans, Mice, NADP metabolism, Nuclear Proteins genetics, Reactive Oxygen Species metabolism, Antioxidants, Nuclear Proteins metabolism, Pentose Phosphate Pathway
- Abstract
TAp73 is a structural homologue of the pre-eminent tumour suppressor p53. However, unlike p53, TAp73 is rarely mutated, and instead is frequently overexpressed in human tumours. It remains unclear whether TAp73 affords an advantage to tumour cells and if so, what the underlying mechanism is. Here we show that TAp73 supports the proliferation of human and mouse tumour cells. TAp73 activates the expression of glucose-6-phosphate dehydrogenase (G6PD), the rate-limiting enzyme of the pentose phosphate pathway (PPP). By stimulating G6PD, TAp73 increases PPP flux and directs glucose to the production of NADPH and ribose, for the synthesis of macromolecules and detoxification of reactive oxygen species (ROS). The growth defect of TAp73-deficient cells can be rescued by either enforced G6PD expression or the presence of nucleosides plus an ROS scavenger. These findings establish a critical role for TAp73 in regulating metabolism, and connect TAp73 and the PPP to oncogenic cell growth.
- Published
- 2013
- Full Text
- View/download PDF
50. PI3K/mTOR inhibition can impair tumor invasion and metastasis in vivo despite a lack of antiproliferative action in vitro: implications for targeted therapy.
- Author
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Wander SA, Zhao D, Besser AH, Hong F, Wei J, Ince TA, Milikowski C, Bishopric NH, Minn AJ, Creighton CJ, and Slingerland JM
- Subjects
- Animals, Bone Neoplasms mortality, Bone Neoplasms secondary, Breast Neoplasms mortality, Breast Neoplasms pathology, Cell Line, Tumor, Cell Movement drug effects, Cell Proliferation drug effects, Cyclin-Dependent Kinase Inhibitor p27 genetics, Cyclin-Dependent Kinase Inhibitor p27 metabolism, Cytoplasm metabolism, Disease-Free Survival, Female, Gene Expression, Gene Knockdown Techniques, Humans, Kaplan-Meier Estimate, Mice, Mice, Inbred BALB C, Mice, Nude, Molecular Targeted Therapy, Neoplasm Invasiveness, RNA, Small Interfering genetics, Signal Transduction drug effects, Tumor Burden drug effects, Xenograft Model Antitumor Assays, Bone Neoplasms prevention & control, Breast Neoplasms drug therapy, Phosphoinositide-3 Kinase Inhibitors, Pyridones pharmacology, Pyrimidines pharmacology, TOR Serine-Threonine Kinases antagonists & inhibitors
- Abstract
Oncogenic PI3K/mTOR activation is frequently observed in human cancers and activates cell motility via p27 phosphorylations at T157 and T198. Here we explored the potential for a novel PI3K/mTOR inhibitor to inhibit tumor invasion and metastasis. An MDA-MB-231 breast cancer line variant, MDA-MB-231-1833, with high metastatic bone tropism, was treated with a novel catalytic PI3K/mTOR inhibitor, PF-04691502, at nM doses that did not impair proliferation. Effects on tumor cell motility, invasion, p27 phosphorylation, localization, and bone metastatic outgrowth were assayed. MDA-MB-231-1833 showed increased PI3K/mTOR activation, high levels of cytoplasmic p27pT157pT198 and increased cell motility and invasion in vitro versus parental. PF-04691502 treatment, at a dose that did not affect proliferation, reduced total and cytoplasmic p27, decreased p27pT157pT198 and restored cell motility and invasion to levels seen in MDA-MB-231. p27 knockdown in MDA-MB-231-1833 phenocopied PI3K/mTOR inhibition, whilst overexpression of the phosphomimetic mutant p27T157DT198D caused resistance to the anti-invasive effects of PF-04691502. Pre-treatment of MDA-MB-231-1833 with PF-04691502 significantly impaired metastatic tumor formation in vivo, despite lack of antiproliferative effects in culture and little effect on primary orthotopic tumor growth. A further link between cytoplasmic p27 and metastasis was provided by a study of primary human breast cancers which showed cytoplasmic p27 is associated with increased lymph nodal metastasis and reduced survival. Novel PI3K/mTOR inhibitors may oppose tumor metastasis independent of their growth inhibitory effects, providing a rationale for clinical investigation of PI3K/mTOR inhibitors in settings to prevent micrometastasis. In primary human breast cancers, cytoplasmic p27 is associated with worse outcomes and increased nodal metastasis, and may prove useful as a marker of both PI3K/mTOR activation and PI3K/mTOR inhibitor efficacy.
- Published
- 2013
- Full Text
- View/download PDF
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