289 results on '"Cleveland JL"'
Search Results
2. Activation of apoptosis associated with enforced myc expression in myeloid progenitor cells is dominant to the suppression of apoptosis by interleukin-3 or erythropoietin
- Author
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Askew, DS, primary, Ihle, JN, additional, and Cleveland, JL, additional
- Published
- 1993
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3. Identification of novel protein tyrosine phosphatases of hematopoietic cells by polymerase chain reaction amplification
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Yi, T, primary, Cleveland, JL, additional, and Ihle, JN, additional
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- 1991
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4. Guidelines for infection control in dental health-care settings -- 2003.
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Kohn WG, Collins AS, Cleveland JL, Harte JA, Eklund KJ, and Malvitz DM
- Abstract
This report consolidates previous recommendations and adds new ones for infection control in dental settings. Recommendations are provided regarding]) educating and protecting dental health-care personnel; 2) preventing transmission of bloodborne pathogens; 3) hand hygiene; 4) personal protective equipment; 5) contact dermatitis and latex hypersensitivity; 6) sterilization and disinfection of patient-care items; 7) environmental infection control; 8) dental unit waterlines, biofilm, and water quality; and 9) special considerations (e.g., dental handpieces and other devices, radiology, parenteral medications, oral surgical procedures, and dental laboratories). These recommendations were developed in collaboration with and after review by authorities on infection control from CDC and other public agencies, academia, and private and professional organizations. [ABSTRACT FROM AUTHOR]
- Published
- 2003
5. Research and professional briefs. Similar glycemic responses to high versus moderate sucrose-containing foods in test meals for adolescents with type 1 diabetes and fasting euglycemia.
- Author
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Rickard KA, Cleveland JL, Loghmani ES, Fineberg NS, and Freidenberg GR
- Published
- 2001
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6. The Many Faces of Hypusinated eIF5A: Cell Context-Specific Effects of the Hypusine Circuit and Implications for Human Health.
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Nakanishi S and Cleveland JL
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- Humans, Animals, Oxidoreductases Acting on CH-NH Group Donors genetics, Oxidoreductases Acting on CH-NH Group Donors metabolism, Mixed Function Oxygenases genetics, Mixed Function Oxygenases metabolism, Protein Biosynthesis, Mice, Eukaryotic Translation Initiation Factor 5A, Peptide Initiation Factors metabolism, Peptide Initiation Factors genetics, RNA-Binding Proteins metabolism, RNA-Binding Proteins genetics, Lysine metabolism, Lysine analogs & derivatives
- Abstract
The unique amino acid hypusine [N
ε -(4-amino-2-hydroxybutyl)lysine] is exclusively formed on the translational regulator eukaryotic initiation factor 5A (eIF5A) via a process coined hypusination. Hypusination is mediated by two enzymes, deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH), and hypusinated eIF5A (eIF5AHyp ) promotes translation elongation by alleviating ribosome pauses at amino acid motifs that cause structural constraints, and it also facilitates translation initiation and termination. Accordingly, eIF5AHyp has diverse biological functions that rely on translational control of its targets. Homozygous deletion of Eif5a , Dhps , or Dohh in mice leads to embryonic lethality, and heterozygous germline variants in EIF5A and biallelic variants in DHPS and DOHH are associated with rare inherited neurodevelopmental disorders, underscoring the importance of the hypusine circuit for embryonic and neuronal development. Given the pleiotropic effects of eIF5AHyp , a detailed understanding of the cell context-specific intrinsic roles of eIF5AHyp and of the chronic versus acute effects of eIF5AHyp inhibition is necessary to develop future strategies for eIF5AHyp -targeted therapy to treat various human health problems. Here, we review the most recent studies documenting the intrinsic roles of eIF5AHyp in different tissues/cell types under normal or pathophysiological conditions and discuss these unique aspects of eIF5AHyp -dependent translational control.- Published
- 2024
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7. Trisomy 8 Defines a Distinct Subtype of Myeloproliferative Neoplasms Driven by the MYC-Alarmin Axis.
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Vincelette ND, Yu X, Kuykendall AT, Moon J, Su S, Cheng CH, Sammut R, Razabdouski TN, Nguyen HV, Eksioglu EA, Chan O, Al Ali N, Patel PC, Lee DH, Nakanishi S, Ferreira RB, Hyjek E, Mo Q, Cory S, Lawrence HR, Zhang L, Murphy DJ, Komrokji RS, Lee D, Kaufmann SH, Cleveland JL, and Yun S
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- Humans, Animals, Mice, Primary Myelofibrosis genetics, Primary Myelofibrosis pathology, Primary Myelofibrosis metabolism, Signal Transduction genetics, Chromosomes, Human, Pair 8 genetics, Trisomy genetics, Proto-Oncogene Proteins c-myc genetics, Proto-Oncogene Proteins c-myc metabolism, Calgranulin B genetics, Calgranulin B metabolism, Myeloproliferative Disorders genetics, Myeloproliferative Disorders metabolism, Myeloproliferative Disorders pathology
- Abstract
Despite advances in understanding the genetic abnormalities in myeloproliferative neoplasms (MPN) and the development of JAK2 inhibitors, there is an urgent need to devise new treatment strategies, particularly for patients with triple-negative (TN) myelofibrosis (MF) who lack mutations in the JAK2 kinase pathway and have very poor clinical outcomes. Here we report that MYC copy number gain and increased MYC expression frequently occur in TN-MF and that MYC-directed activation of S100A9, an alarmin protein that plays pivotal roles in inflammation and innate immunity, is necessary and sufficient to drive development and progression of MF. Notably, the MYC-S100A9 circuit provokes a complex network of inflammatory signaling that involves numerous hematopoietic cell types in the bone marrow microenvironment. Accordingly, genetic ablation of S100A9 or treatment with small molecules targeting the MYC-S100A9 pathway effectively ameliorates MF phenotypes, highlighting the MYC-alarmin axis as a novel therapeutic vulnerability for this subgroup of MPNs. Significance: This study establishes that MYC expression is increased in TN-MPNs via trisomy 8, that a MYC-S100A9 circuit manifest in these cases is sufficient to provoke myelofibrosis and inflammation in diverse hematopoietic cell types in the BM niche, and that the MYC-S100A9 circuit is targetable in TN-MPNs., (©2024 American Association for Cancer Research.)
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- 2024
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8. Lung adenocarcinomas without driver genes converge to common adaptive strategies through diverse genetic, epigenetic, and niche construction evolutionary pathways.
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Gatenby RA, Luddy KA, Teer JK, Berglund A, Freischel AR, Carr RM, Lam AE, Pienta KJ, Amend SR, Austin RH, Hammarlund EU, Cleveland JL, Tsai KY, and Brown JS
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- Humans, Gene Expression Regulation, Neoplastic genetics, Evolution, Molecular, Tumor Microenvironment genetics, Lung Neoplasms genetics, Lung Neoplasms pathology, Adenocarcinoma of Lung genetics, Adenocarcinoma of Lung pathology, Mutation, Epigenesis, Genetic genetics
- Abstract
Somatic evolution selects cancer cell phenotypes that maximize survival and proliferation in dynamic environments. Although cancer cells are molecularly heterogeneous, we hypothesized convergent adaptive strategies to common host selection forces can be inferred from patterns of epigenetic and genetic evolutionary selection in similar tumors. We systematically investigated gene mutations and expression changes in lung adenocarcinomas with no common driver genes (n = 313). Although 13,461 genes were mutated in at least one sample, only 376 non-synonymous mutations evidenced positive evolutionary selection with conservation of 224 genes, while 1736 and 2430 genes exhibited ≥ two-fold increased and ≥ 50% decreased expression, respectively. Mutations under positive selection are more frequent in genes with significantly altered expression suggesting they often "hardwire" pre-existing epigenetically driven adaptations. Conserved genes averaged 16-fold higher expression in normal lung tissue compared to those with selected mutations demonstrating pathways necessary for both normal cell function and optimal cancer cell fitness. The convergent LUAD phenotype exhibits loss of differentiated functions and cell-cell interactions governing tissue organization. Conservation with increased expression is found in genes associated with cell cycle, DNA repair, p53 pathway, epigenetic modifiers, and glucose metabolism. No canonical driver gene pathways exhibit strong positive selection, but extensive down-regulation of membrane ion channels suggests decreased transmembrane potential may generate persistent proliferative signals. NCD LUADs perform niche construction generating a stiff, immunosuppressive microenvironment through selection of specific collagens and proteases. NCD LUADs evolve to a convergent phenotype through a network of interconnected genetic, epigenetic, and ecological pathways., (© 2024. The Author(s).)
- Published
- 2024
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9. Transcriptional Plasticity Drives IMiD and p300 Inhibitor Resistance in Multiple Myeloma.
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Yun S and Cleveland JL
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- Humans, Nuclear Proteins metabolism, Immunomodulating Agents, Peptide Hydrolases genetics, Peptide Hydrolases therapeutic use, Bromodomain Containing Proteins, Transcription Factors genetics, Cell Cycle Proteins, Multiple Myeloma drug therapy, Multiple Myeloma genetics
- Abstract
Summary: In this issue of Blood Cancer Discovery, Neri, Barwick, and colleagues and Welsh, Barwick, and colleagues performed RNA sequencing, chromatin immunoprecipitation sequencing, assay for transposase-accessible chromatin using sequencing, and genetic studies to characterize the underlying mechanisms of immunomodulatory drug (IMiD) resistance in multiple myeloma. They demonstrated that IMiD resistance is driven by sustained expression of MYC and IRF4 via transcriptional plasticity that involves induction of ETV4 and BATF proteins, the binding of these proteins to their super-enhancers, and the recruitment of BRD4 and p300. Finally, these studies suggest IMiD and p300 inhibitor combination as a promising therapeutic strategy in multiple myeloma. See related article by Neri, Barwick, et al., p. 56 (9). See related article by Welsh, Barwick, et al., p. 34 (10)., (©2023 American Association for Cancer Research.)
- Published
- 2024
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10. CK1δ and CK1ε Signaling Sustains Mitochondrial Metabolism and Cell Survival in Multiple Myeloma.
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Burger KL, Fernandez MR, Meads MB, Sudalagunta P, Oliveira PS, Renatino Canevarolo R, Alugubelli RR, Tungsevik A, De Avila G, Silva M, Graeter AI, Dai HA, Vincelette ND, Prabhu A, Magaletti D, Yang C, Li W, Kulkarni A, Hampton O, Koomen JM, Roush WR, Monastyrskyi A, Berglund AE, Silva AS, Cleveland JL, and Shain KH
- Subjects
- Humans, Cell Survival, Phosphorylation, Disease Progression, Casein Kinase Idelta genetics, Casein Kinase Idelta metabolism, Multiple Myeloma genetics
- Abstract
Multiple myeloma remains an incurable malignancy due to acquisition of intrinsic programs that drive therapy resistance. Here we report that casein kinase-1δ (CK1δ) and CK1ε are therapeutic targets in multiple myeloma that are necessary to sustain mitochondrial metabolism. Specifically, the dual CK1δ/CK1ε inhibitor SR-3029 had potent in vivo and ex vivo anti-multiple myeloma activity, including against primary multiple myeloma patient specimens. RNA sequencing (RNA-seq) and metabolic analyses revealed inhibiting CK1δ/CK1ε disables multiple myeloma metabolism by suppressing genes involved in oxidative phosphorylation (OxPhos), reducing citric acid cycle intermediates, and suppressing complexes I and IV of the electron transport chain. Finally, sensitivity of multiple myeloma patient specimens to SR-3029 correlated with elevated expression of mitochondrial genes, and RNA-seq from 687 multiple myeloma patient samples revealed that increased CSNK1D, CSNK1E, and OxPhos genes correlate with disease progression and inferior outcomes. Thus, increases in mitochondrial metabolism are a hallmark of multiple myeloma progression that can be disabled by targeting CK1δ/CK1ε., Significance: CK1δ and CK1ε are attractive therapeutic targets in multiple myeloma whose expression increases with disease progression and connote poor outcomes, and that are necessary to sustain expression of genes directing OxPhos., (©2023 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2023
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11. AACR Cancer Centers Alliance: Fostering Collaboration and Innovation to Advance Lifesaving Scientific Discoveries for Patients.
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Arteaga CL, Cleveland JL, Foti M, Mesa RA, Weiner LM, Willman CL, and Tuveson DA
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- Humans, United States, Delivery of Health Care, Health Policy, Public Health, Neoplasms therapy
- Abstract
Summary: Basic and clinical cancer research discoveries stemming from the nation's cancer centers have markedly improved outcomes for many cancer patients. Despite this forward momentum in our progress against this complex disease, cancer in all its forms remains a major public health challenge that touches the lives of nearly every American, either directly or indirectly. The newly formed AACR Cancer Centers Alliance will accelerate the pace of discovery by providing an ongoing mechanism for transferring new knowledge, sharing resources, developing national demonstration projects, and driving innovation that impacts cancer science, cancer care delivery, and science and health policy., (©2023 American Association for Cancer Research.)
- Published
- 2023
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12. Modulating the polyamine/hypusine axis controls generation of CD8+ tissue-resident memory T cells.
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Elmarsafawi AG, Hesterberg RS, Fernandez MR, Yang C, Darville LN, Liu M, Koomen JM, Phanstiel O 4th, Atkins R, Mullinax JE, Pilon-Thomas SA, Locke FL, Epling-Burnette PK, and Cleveland JL
- Subjects
- Humans, Memory T Cells, Glutamine metabolism, CD8-Positive T-Lymphocytes metabolism, Polyamines metabolism, Spermidine metabolism
- Abstract
Glutaminolysis is a hallmark of the activation and metabolic reprogramming of T cells. Isotopic tracer analyses of antigen-activated effector CD8+ T cells revealed that glutamine is the principal carbon source for the biosynthesis of polyamines putrescine, spermidine, and spermine. These metabolites play critical roles in activation-induced T cell proliferation, as well as for the production of hypusine, which is derived from spermidine and is covalently linked to the translation elongation factor eukaryotic translation initiation factor 5A (eIF5A). Here, we demonstrated that the glutamine/polyamine/hypusine axis controlled the expression of CD69, an important regulator of tissue-resident memory T cells (Trm). Inhibition of this circuit augmented the development of Trm cells ex vivo and in vivo in the BM, a well-established niche for Trm cells. Furthermore, blocking the polyamine/hypusine axis augmented CD69 expression as well as IFN-γ and TNF-α production in (a) human CD8+ T cells from peripheral blood and sarcoma tumor infiltrating lymphocytes and (b) human CD8+ CAR-T cells. Collectively, these findings support the notion that the polyamine-hypusine circuit can be exploited to modulate Trm cells for therapeutic benefit.
- Published
- 2023
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13. Associations between prediagnostic aspirin use and ovarian tumor gene expression.
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Sasamoto N, Stewart PA, Wang T, Thompson ZJ, Yoder SJ, Hecht JL, Cleveland JL, Conejo-Garcia J, Fridley BL, Terry KL, and Tworoger SS
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- Female, Humans, Case-Control Studies, Gene Expression, Estrogens, Aspirin adverse effects, Ovarian Neoplasms pathology
- Abstract
Background: Aspirin use has been associated with reduced ovarian cancer risk, yet the underlying biological mechanisms are not fully understood. To gain mechanistic insights, we assessed the association between prediagnosis low and regular-dose aspirin use and gene expression profiles in ovarian tumors., Methods: RNA sequencing was performed on high-grade serous, poorly differentiated, and high-grade endometrioid ovarian cancer tumors from the Nurses' Health Study (NHS), NHSII, and New England Case-Control Study (n = 92 cases for low, 153 cases for regular-dose aspirin). Linear regression identified differentially expressed genes associated with aspirin use, adjusted for birth decade and cohort. False discovery rates (FDR) were used to account for multiple testing and gene set enrichment analysis was used to identify biological pathways., Results: No individual genes were significantly differentially expressed in ovarian tumors in low or regular-dose aspirin users accounting for multiple comparisons. However, current versus never use of low-dose aspirin was associated with upregulation of immune pathways (e.g., allograft rejection, FDR = 5.8 × 10
-10 ; interferon-gamma response, FDR = 2.0 × 10-4 ) and downregulation of estrogen response pathways (e.g., estrogen response late, FDR = 4.9 × 10-8 ). Ovarian tumors from current regular aspirin users versus never users were also associated with upregulation in interferon pathways (FDR <1.5 × 10-4 ) and downregulation of multiple extracellular matrix (ECM) architecture pathways (e.g., ECM organization, 4.7 × 10-8 )., Conclusion: Our results suggest low and regular-dose aspirin may impair ovarian tumorigenesis in part via enhancing adaptive immune response and decreasing metastatic potential supporting the likely differential effects on ovarian carcinogenesis and progression by dose of aspirin., (© 2023 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)- Published
- 2023
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14. The Polyamine-Hypusine Circuit Controls an Oncogenic Translational Program Essential for Malignant Conversion in MYC-Driven Lymphoma.
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Nakanishi S, Li J, Berglund AE, Kim Y, Zhang Y, Zhang L, Yang C, Song J, Mirmira RG, and Cleveland JL
- Subjects
- Humans, Polyamines metabolism, Proteomics, Neoplasms, Lymphoma
- Abstract
The MYC oncoprotein is activated in a broad spectrum of human malignancies and transcriptionally reprograms the genome to drive cancer cell growth. Given this, it is unclear if targeting a single effector of MYC will have therapeutic benefit. MYC activates the polyamine-hypusine circuit, which posttranslationally modifies the eukaryotic translation factor eIF5A. The roles of this circuit in cancer are unclear. Here we report essential intrinsic roles for hypusinated eIF5A in the development and maintenance of MYC-driven lymphoma, where the loss of eIF5A hypusination abolishes malignant transformation of MYC-overexpressing B cells. Mechanistically, integrating RNA sequencing, ribosome sequencing, and proteomic analyses revealed that efficient translation of select targets is dependent upon eIF5A hypusination, including regulators of G1-S phase cell-cycle progression and DNA replication. This circuit thus controls MYC's proliferative response, and it is also activated across multiple malignancies. These findings suggest the hypusine circuit as a therapeutic target for several human tumor types., Significance: Elevated EIF5A and the polyamine-hypusine circuit are manifest in many malignancies, including MYC-driven tumors, and eIF5A hypusination is necessary for MYC proliferative signaling. Not-ably, this circuit controls an oncogenic translational program essential for the development and maintenance of MYC-driven lymphoma, supporting this axis as a target for cancer prevention and treatment. See related commentary by Wilson and Klein, p. 248. This article is highlighted in the In This Issue feature, p. 247., (©2023 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2023
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15. Programming inactive RNA-binding small molecules into bioactive degraders.
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Tong Y, Lee Y, Liu X, Childs-Disney JL, Suresh BM, Benhamou RI, Yang C, Li W, Costales MG, Haniff HS, Sievers S, Abegg D, Wegner T, Paulisch TO, Lekah E, Grefe M, Crynen G, Van Meter M, Wang T, Gibaut QMR, Cleveland JL, Adibekian A, Glorius F, Waldmann H, and Disney MD
- Subjects
- Humans, Genes, jun genetics, Genes, myc genetics, Nucleic Acid Conformation, Structure-Activity Relationship, Substrate Specificity, Transcriptome, MicroRNAs antagonists & inhibitors, MicroRNAs chemistry, MicroRNAs genetics, MicroRNAs metabolism, RNA, Messenger antagonists & inhibitors, RNA, Messenger chemistry, RNA, Messenger genetics, RNA, Messenger metabolism, Endoribonucleases chemistry, Endoribonucleases metabolism
- Abstract
Target occupancy is often insufficient to elicit biological activity, particularly for RNA, compounded by the longstanding challenges surrounding the molecular recognition of RNA structures by small molecules. Here we studied molecular recognition patterns between a natural-product-inspired small-molecule collection and three-dimensionally folded RNA structures. Mapping these interaction landscapes across the human transcriptome defined structure-activity relationships. Although RNA-binding compounds that bind to functional sites were expected to elicit a biological response, most identified interactions were predicted to be biologically inert as they bind elsewhere. We reasoned that, for such cases, an alternative strategy to modulate RNA biology is to cleave the target through a ribonuclease-targeting chimera, where an RNA-binding molecule is appended to a heterocycle that binds to and locally activates RNase L
1 . Overlay of the substrate specificity for RNase L with the binding landscape of small molecules revealed many favourable candidate binders that might be bioactive when converted into degraders. We provide a proof of concept, designing selective degraders for the precursor to the disease-associated microRNA-155 (pre-miR-155), JUN mRNA and MYC mRNA. Thus, small-molecule RNA-targeted degradation can be leveraged to convert strong, yet inactive, binding interactions into potent and specific modulators of RNA function., (© 2023. The Author(s).)- Published
- 2023
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16. An integrated pre-clerkship curriculum to build cognitive medical schema: It's not just about the content.
- Author
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LeClair RJ, Cleveland JL, Eden K, and Binks AP
- Abstract
Both physiology and pathophysiology are essential disciplines in health professional education however, clinicians do not use this knowledge in isolation. Instead, physicians use inter-disciplinary concepts embedded within integrated cognitive schema (illness scripts) established through experience/knowledge that manifest as expert-level thinking. Our goal was to develop a pre-clerkship curriculum devoid of disciplinary boundaries (akin to the physician's illness script) and enhance learners' clerkship and early clinical performance. As well as developing curricular content, the model considered non-content design elements such as learner characteristics and values, faculty and resources and the impact of curricular and pedagogical changes. The goals of the trans-disciplinary integration were to develop deep learning behaviors through, 1) developing of integrated, cognitive schema to support the transition to expert-level thinking, 2) authentic, contextualization to promote knowledge transfer to the clinical realm 3) allowing autonomous, independent learning, and 4) harnessing the benefits of social learning. The final curricular model was a case-based approach with independent learning of basic concepts, differential diagnosis and illness scripting writing, and concept mapping. Small-group classroom sessions were team-taught with basic scientists and physicians facilitating learners' self-reflection and development of clinical reasoning. Specifications grading was used to assess the products (written illness scripts and concept maps) as well as process (group dynamics) while allowing a greater degree of learner autonomy. Although the model we adopted could be transferred to other program settings, we suggest it is critical to consider both content and non-content elements that are specific to the environment and learner., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 LeClair, Cleveland, Eden and Binks.)
- Published
- 2023
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17. Evolutionary Analysis of TCGA Data Using Over- and Under- Mutated Genes Identify Key Molecular Pathways and Cellular Functions in Lung Cancer Subtypes.
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Freischel AR, Teer JK, Luddy K, Cunningham J, Artzy-Randrup Y, Epstein T, Tsai KY, Berglund A, Cleveland JL, Gillies RJ, Brown JS, and Gatenby RA
- Abstract
We identify critical conserved and mutated genes through a theoretical model linking a gene’s fitness contribution to its observed mutational frequency in a clinical cohort. “Passenger” gene mutations do not alter fitness and have mutational frequencies determined by gene size and the mutation rate. Driver mutations, which increase fitness (and proliferation), are observed more frequently than expected. Non-synonymous mutations in essential genes reduce fitness and are eliminated by natural selection resulting in lower prevalence than expected. We apply this “evolutionary triage” principle to TCGA data from EGFR-mutant, KRAS-mutant, and NEK (non-EGFR/KRAS) lung adenocarcinomas. We find frequent overlap of evolutionarily selected non-synonymous gene mutations among the subtypes suggesting enrichment for adaptations to common local tissue selection forces. Overlap of conserved genes in the LUAD subtypes is rare suggesting negative evolutionary selection is strongly dependent on initiating mutational events during carcinogenesis. Highly expressed genes are more likely to be conserved and significant changes in expression (>20% increased/decreased) are common in genes with evolutionarily selected mutations but not in conserved genes. EGFR-mut cancers have fewer average mutations (89) than KRAS-mut (228) and NEK (313). Subtype-specific variation in conserved and mutated genes identify critical molecular components in cell signaling, extracellular matrix remodeling, and membrane transporters. These findings demonstrate subtype-specific patterns of co-adaptations between the defining driver mutation and somatically conserved genes as well as novel insights into epigenetic versus genetic contributions to cancer evolution.
- Published
- 2022
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18. Ornithine decarboxylase supports ILC3 responses in infectious and autoimmune colitis through positive regulation of IL-22 transcription.
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Peng V, Cao S, Trsan T, Bando JK, Avila-Pacheco J, Cleveland JL, Clish C, Xavier RJ, and Colonna M
- Subjects
- Mice, Animals, Nuclear Receptor Subfamily 1, Group F, Member 3, Interleukin-17, Ornithine Decarboxylase genetics, Immunity, Innate, Putrescine, Th17 Cells metabolism, Ornithine, Anti-Bacterial Agents, Interleukin-22, Colitis genetics, Enterobacteriaceae Infections genetics
- Abstract
Group 3 innate lymphoid cells (ILC3s) are RORγT
+ lymphocytes that are predominately enriched in mucosal tissues and produce IL-22 and IL-17A. They are the innate counterparts of Th17 cells. While Th17 lymphocytes utilize unique metabolic pathways in their differentiation program, it is unknown whether ILC3s make similar metabolic adaptations. We employed single-cell RNA sequencing and metabolomic profiling of intestinal ILC subsets to identify an enrichment of polyamine biosynthesis in ILC3s, converging on the rate-limiting enzyme ornithine decarboxylase (ODC1). In vitro and in vivo studies demonstrated that exogenous supplementation with the polyamine putrescine or its biosynthetic substrate, ornithine, enhanced ILC3 production of IL-22. Conditional deletion of ODC1 in ILC3s impaired mouse antibacterial defense against Citrobacter rodentium infection, which was associated with a decrease in anti-microbial peptide production by the intestinal epithelium. Furthermore, in a model of anti-CD40 colitis, deficiency of ODC1 in ILC3s markedly reduced the production of IL-22 and severity of inflammatory colitis. We conclude that ILC3-intrinsic polyamine biosynthesis facilitates efficient defense against enteric pathogens as well as exacerbates autoimmune colitis, thus representing an attractive target to modulate ILC3 function in intestinal disease.- Published
- 2022
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19. TCR-Independent Metabolic Reprogramming Precedes Lymphoma-Driven Changes in T-cell Fate.
- Author
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Hesterberg RS, Liu M, Elmarsafawi AG, Koomen JM, Welsh EA, Hesterberg SG, Ranatunga S, Yang C, Li W, Lawrence HR, Rodriguez PC, Berglund AE, and Cleveland JL
- Subjects
- Animals, CD4-Positive T-Lymphocytes, Cell Differentiation, Glucose metabolism, Mice, Mice, Transgenic, Receptors, Antigen, T-Cell metabolism, Tumor Microenvironment, Lymphoma metabolism, Lymphoma, B-Cell metabolism
- Abstract
Chronic T-cell receptor (TCR) signaling in the tumor microenvironment is known to promote T-cell dysfunction. However, we reasoned that poorly immunogenic tumors may also compromise T cells by impairing their metabolism. To address this, we assessed temporal changes in T-cell metabolism, fate, and function in models of B-cell lymphoma driven by Myc, a promoter of energetics and repressor of immunogenicity. Increases in lymphoma burden most significantly impaired CD4+ T-cell function and promoted regulatory T cell (Treg) and Th1-cell differentiation. Metabolomic analyses revealed early reprogramming of CD4+ T-cell metabolism, reduced glucose uptake, and impaired mitochondrial function, which preceded changes in T-cell fate. In contrast, B-cell lymphoma metabolism remained robust during tumor progression. Finally, mitochondrial functions were impaired in CD4+ and CD8+ T cells in lymphoma-transplanted OT-II and OT-I transgenic mice, respectively. These findings support a model, whereby early, TCR-independent, metabolic interactions with developing lymphomas limits T cell-mediated immune surveillance., (©2022 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2022
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20. The Radiosensitivity Index Gene Signature Identifies Distinct Tumor Immune Microenvironment Characteristics Associated With Susceptibility to Radiation Therapy.
- Author
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Grass GD, Alfonso JCL, Welsh E, Ahmed KA, Teer JK, Pilon-Thomas S, Harrison LB, Cleveland JL, Mulé JJ, Eschrich SA, Enderling H, and Torres-Roca JF
- Subjects
- Biomarkers, Tumor genetics, Gene Expression Regulation, Neoplastic, Humans, Male, Prognosis, Radiation Tolerance genetics, Transcriptome, Tumor Microenvironment genetics, CD8-Positive T-Lymphocytes, Neoplasms genetics, Neoplasms radiotherapy
- Abstract
Purpose: Radiation therapy (RT) is a mainstay of cancer care, and accumulating evidence suggests the potential for synergism with components of the immune response. However, few data describe the tumor immune contexture in relation to RT sensitivity. To address this challenge, we used the radiation sensitivity index (RSI) gene signature to estimate the RT sensitivity of >10,000 primary tumors and characterized their immune microenvironments in relation to the RSI., Methods and Materials: We analyzed gene expression profiles of 10,469 primary tumors (31 types) within a prospective tissue collection protocol. The RT sensitivity of each tumor was estimated by the RSI and respective distributions were characterized. The tumor biology measured by the RSI was evaluated by differentially expressed genes combined with single sample gene set enrichment analysis. Differences in the expression of immune regulatory molecules were assessed and deconvolution algorithms were used to estimate immune cell infiltrates in relation to the RSI. A subset (n = 2368) of tumors underwent DNA sequencing for mutational frequency characterization., Results: We identified a wide range of RSI values within and across various tumor types, with several demonstrating nonunimodal distributions (eg, colon, renal, lung, prostate, esophagus, pancreas, and PAM50 breast subtypes; P < .05). Across all tumor types, stratifying RSI at a tumor type-specific median identified 7148 differentially expressed genes, of which 146 were coordinate in direction. Network topology analysis demonstrates RSI measures a coordinated STAT1, IRF1, and CCL4/MIP-1β transcriptional network. Tumors with an estimated high sensitivity to RT demonstrated distinct enrichment of interferon-associated signaling pathways and immune cell infiltrates (eg, CD8
+ T cells, activated natural killer cells, M1-macrophages; q < 0.05), which was in the context of diverse expression patterns of various immunoregulatory molecules., Conclusions: This analysis describes the immune microenvironments of patient tumors in relation to the RSI gene expression signature., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
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21. Immunodepletion of MDSC by AMV564, a novel bivalent, bispecific CD33/CD3 T cell engager, ex vivo in MDS and melanoma.
- Author
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Cheng P, Chen X, Dalton R, Calescibetta A, So T, Gilvary D, Ward G, Smith V, Eckard S, Fox JA, Guenot J, Markowitz J, Cleveland JL, Wright KL, List AF, Wei S, and Eksioglu EA
- Subjects
- Animals, Humans, Sialic Acid Binding Ig-like Lectin 3, T-Lymphocytes, Antibodies, Bispecific pharmacology, Antineoplastic Agents pharmacology, Melanoma drug therapy, Myelodysplastic Syndromes drug therapy, Myeloid-Derived Suppressor Cells
- Abstract
We have reported previously that CD33
hi myeloid-derived suppressor cells (MDSCs) play a direct role in the pathogenesis of myelodysplastic syndromes (MDSs) and that their sustained activation contributes to hematopoietic and immune impairment, including modulation of PD1/PDL1. MDSCs can also limit the clinical activity of immune checkpoint inhibition in solid malignancies. We hypothesized that depletion of MDSCs may ameliorate resistance to checkpoint inhibitors and, hence, targeted them with AMV564 combined with anti-PD1 in MDS bone marrow (BM) mononuclear cells (MNCs) enhanced activation of cytotoxic T cells. AMV564 was active in vivo in a leukemia xenograft model when co-administered with healthy donor peripheral blood MNCs (PBMCs). Our findings provide a strong rationale for clinical investigation of AMV564 as a single agent or in combination with an anti-PD1 antibody and in particular for treatment of cancers resistant to checkpoint inhibitors., Competing Interests: Declaration of interests S.E. and V.S. are employees of Amphivena Therapeutics, Inc. S.W. and A.L. are scientific/medical advisors for Amphivena Therapeutics, Inc. The S.W. and A.F.L. laboratories received unrelated support from Celgene and Genentech. A.F.L. has provided consulting services and has received research funding support from multiple other pharmaceutical and biotechnology companies that pursue cancer research, including Celgene, Precision BioSciences, Prelude Therapeutics, and CTI Biopharma, and serves on the scientific advisory boards of Aileron and CBMG. A.F.L. also has potential royalty income from IP he developed related to cancer treatment and is a member of the MDS Foundation board of directors. S.W. has received unrelated support from Blackbird BioFinance, LLC. J.A.F. is a consultant for and owns Amphivena stocks and is currently employed by Sunesis Pharmaceuticals., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2022
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22. Disrupting the MYC-TFEB Circuit Impairs Amino Acid Homeostasis and Provokes Metabolic Anergy.
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Fernandez MR, Schaub FX, Yang C, Li W, Yun S, Schaub SK, Dorsey FC, Liu M, Steeves MA, Ballabio A, Tzankov A, Chen Z, Koomen JM, Berglund AE, and Cleveland JL
- Subjects
- Amino Acids metabolism, Animals, Autophagy physiology, Homeostasis, Humans, Mice, Proteasome Endopeptidase Complex metabolism, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, Lysosomes metabolism, Proto-Oncogene Proteins c-myc genetics
- Abstract
MYC family oncoproteins are regulators of metabolic reprogramming that sustains cancer cell anabolism. Normal cells adapt to nutrient-limiting conditions by activating autophagy, which is required for amino acid (AA) homeostasis. Here we report that the autophagy pathway is suppressed by Myc in normal B cells, in premalignant and neoplastic B cells of Eμ-Myc transgenic mice, and in human MYC-driven Burkitt lymphoma. Myc suppresses autophagy by antagonizing the expression and function of transcription factor EB (TFEB), a master regulator of autophagy. Mechanisms that sustained AA pools in MYC-expressing B cells include coordinated induction of the proteasome and increases in AA transport. Reactivation of the autophagy-lysosomal pathway by TFEB disabled the malignant state by disrupting mitochondrial functions, proteasome activity, AA transport, and AA and nucleotide metabolism, leading to metabolic anergy, growth arrest, and apoptosis. This phenotype provides therapeutic opportunities to disable MYC-driven malignancies, including AA restriction and treatment with proteasome inhibitors., Significance: MYC suppresses TFEB and autophagy and controls amino acid homeostasis by upregulating amino acid transport and the proteasome, and reactivation of TFEB disables the metabolism of MYC-driven tumors., (©2022 The Authors; Published by the American Association for Cancer Research.)
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- 2022
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23. Novel Transcriptomic Interactions Between Immune Content and Genomic Classifier Predict Lethal Outcomes in High-grade Prostate Cancer.
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Yamoah K, Awasthi S, Mahal BA, Zhao SG, Grass GD, Berglund A, Abraham-Miranda J, Gerke T, Rounbehler RJ, Davicioni E, Liu Y, Park J, Cleveland JL, Pow-Sang JM, Fernandez D, Torres-Roca J, Karnes RJ, Schaeffer E, Freedland SJ, Spratt DE, Den RB, Rebbeck TR, and Feng F
- Subjects
- Humans, Male, Neoplasm Grading, Prostate pathology, Prostatectomy adverse effects, Prostatic Neoplasms genetics, Prostatic Neoplasms pathology, Prostatic Neoplasms therapy, Transcriptome
- Abstract
Grade group 4 and 5 (GG-45) prostate cancer (PCa) patients are at the highest risk of lethal outcomes, yet lack genomic risk stratification for prognosis and treatment selection. Here, we assess whether transcriptomic interactions between tumor immune content score (ICS) and the Decipher genomic classifier can identify most lethal subsets of GG-45 PCa. We utilized whole transcriptome data from 8071 tumor tissue (6071 prostatectomy and 2000 treatment-naïve biopsy samples) to derive four immunogenomic subtypes using ICS and Decipher. When compared across all grade groups, GG-45 samples had the highest proportion of most aggressive subtype-ICS
High /DecipherHigh . Subsequent analyses within the GG-45 patient samples (n = 1420) revealed that the ICSHigh /DecipherHigh subtype was associated with increased genomic radiosensitivity. Additionally, in a multivariable model (n = 335), ICSHigh /DecipherHigh subtype had a significantly higher risk of distant metastasis (hazard ratio [HR] = 5.41; 95% confidence interval [CI], 2.76-10.6; p ≤ 0.0001) and PCa-specific mortality (HR = 10.6; 95% CI, 4.18-26.94; p ≤ 0.0001) as compared with ICSLow /DecipherLow . The novel immunogenomic subtypes establish a very strong synergistic interaction between ICS and Decipher in identifying GG-45 patients who experience the most lethal outcomes. PATIENT SUMMARY: In this analysis, we identified a novel interaction between the total immune content of prostate tumors and genomic classifier to identify the most lethal subset of patients with grade groups 4 and 5. Our results will aid in the subtyping of aggressive prostate cancer patients who may benefit from combined immune-radiotherapy modalities., (Copyright © 2020 European Association of Urology. Published by Elsevier B.V. All rights reserved.)- Published
- 2022
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24. Tumor-immune ecosystem dynamics define an individual Radiation Immune Score to predict pan-cancer radiocurability.
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Alfonso JCL, Grass GD, Welsh E, Ahmed KA, Teer JK, Pilon-Thomas S, Harrison LB, Cleveland JL, Mulé JJ, Eschrich SA, Torres-Roca JF, and Enderling H
- Subjects
- Humans, Lung Neoplasms genetics, Lung Neoplasms immunology, Lung Neoplasms radiotherapy, Prognosis, Radiation Tolerance immunology, Radiotherapy, Survival Rate, Biomarkers metabolism, Gene Expression Regulation, Neoplastic, Lung Neoplasms pathology, Lymphocytes, Tumor-Infiltrating immunology, Radiation Tolerance genetics, Tumor Microenvironment
- Abstract
Radiotherapy efficacy is the result of radiation-mediated cytotoxicity coupled with stimulation of antitumor immune responses. We develop an in silico 3-dimensional agent-based model of diverse tumor-immune ecosystems (TIES) represented as anti- or pro-tumor immune phenotypes. We validate the model in 10,469 patients across 31 tumor types by demonstrating that clinically detected tumors have pro-tumor TIES. We then quantify the likelihood radiation induces antitumor TIES shifts toward immune-mediated tumor elimination by developing the individual Radiation Immune Score (iRIS). We show iRIS distribution across 31 tumor types is consistent with the clinical effectiveness of radiotherapy, and in combination with a molecular radiosensitivity index (RSI) combines to predict pan-cancer radiocurability. We show that iRIS correlates with local control and survival in a separate cohort of 59 lung cancer patients treated with radiation. In combination, iRIS and RSI predict radiation-induced TIES shifts in individual patients and identify candidates for radiation de-escalation and treatment escalation. This is the first clinically and biologically validated computational model to simulate and predict pan-cancer response and outcomes via the perturbation of the TIES by radiotherapy., (Copyright © 2021. Published by Elsevier Inc.)
- Published
- 2021
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25. Polyamine Homeostasis in Development and Disease.
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Nakanishi S and Cleveland JL
- Subjects
- Cell Proliferation, Homeostasis, Humans, Neoplasms drug therapy, Polyamines metabolism
- Abstract
Polycationic polyamines are present in nearly all living organisms and are essential for mammalian cell growth and survival, and for development. These positively charged molecules are involved in a variety of essential biological processes, yet their underlying mechanisms of action are not fully understood. Several studies have shown both beneficial and detrimental effects of polyamines on human health. In cancer, polyamine metabolism is frequently dysregulated, and elevated polyamines have been shown to promote tumor growth and progression, suggesting that targeting polyamines is an attractive strategy for therapeutic intervention. In contrast, polyamines have also been shown to play critical roles in lifespan, cardiac health and in the development and function of the brain. Accordingly, a detailed understanding of mechanisms that control polyamine homeostasis in human health and disease is needed to develop safe and effective strategies for polyamine-targeted therapy.
- Published
- 2021
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26. KLK3 SNP-SNP interactions for prediction of prostate cancer aggressiveness.
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Lin HY, Huang PY, Cheng CH, Tung HY, Fang Z, Berglund AE, Chen A, French-Kwawu J, Harris D, Pow-Sang J, Yamoah K, Cleveland JL, Awasthi S, Rounbehler RJ, Gerke T, Dhillon J, Eeles R, Kote-Jarai Z, Muir K, Schleutker J, Pashayan N, Neal DE, Nielsen SF, Nordestgaard BG, Gronberg H, Wiklund F, Giles GG, Haiman CA, Travis RC, Stanford JL, Kibel AS, Cybulski C, Khaw KT, Maier C, Thibodeau SN, Teixeira MR, Cannon-Albright L, Brenner H, Kaneva R, Pandha H, Srinivasan S, Clements J, Batra J, and Park JY
- Subjects
- Biomarkers, Tumor genetics, Epistasis, Genetic, Genetic Predisposition to Disease, Genotype, Humans, Male, Polymorphism, Single Nucleotide, Prostatic Neoplasms pathology, Kallikreins genetics, Prostate-Specific Antigen genetics, Prostatic Neoplasms genetics
- Abstract
Risk classification for prostate cancer (PCa) aggressiveness and underlying mechanisms remain inadequate. Interactions between single nucleotide polymorphisms (SNPs) may provide a solution to fill these gaps. To identify SNP-SNP interactions in the four pathways (the angiogenesis-, mitochondria-, miRNA-, and androgen metabolism-related pathways) associated with PCa aggressiveness, we tested 8587 SNPs for 20,729 cases from the PCa consortium. We identified 3 KLK3 SNPs, and 1083 (P < 3.5 × 10
-9 ) and 3145 (P < 1 × 10-5 ) SNP-SNP interaction pairs significantly associated with PCa aggressiveness. These SNP pairs associated with PCa aggressiveness were more significant than each of their constituent SNP individual effects. The majority (98.6%) of the 3145 pairs involved KLK3. The 3 most common gene-gene interactions were KLK3-COL4A1:COL4A2, KLK3-CDH13, and KLK3-TGFBR3. Predictions from the SNP interaction-based polygenic risk score based on 24 SNP pairs are promising. The prevalence of PCa aggressiveness was 49.8%, 21.9%, and 7.0% for the PCa cases from our cohort with the top 1%, middle 50%, and bottom 1% risk profiles. Potential biological functions of the identified KLK3 SNP-SNP interactions were supported by gene expression and protein-protein interaction results. Our findings suggest KLK3 SNP interactions may play an important role in PCa aggressiveness.- Published
- 2021
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27. Transcriptional programming drives Ibrutinib-resistance evolution in mantle cell lymphoma.
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Zhao X, Wang MY, Jiang H, Lwin T, Park PM, Gao J, Meads MB, Ren Y, Li T, Sun J, Fahmi NA, Singh S, Sehgal L, Wang X, Silva AS, Sotomayor EM, Shain KH, Cleveland JL, Wang M, Zhang W, Qi J, Shah BD, and Tao J
- Subjects
- Adenine pharmacology, Adenine therapeutic use, Animals, Cell Cycle Proteins antagonists & inhibitors, Cell Cycle Proteins metabolism, Cell Line, Tumor, Cyclin-Dependent Kinase 9 antagonists & inhibitors, Cyclin-Dependent Kinase 9 metabolism, Drug Resistance, Neoplasm drug effects, Enhancer Elements, Genetic genetics, Humans, Lymphoma, Mantle-Cell enzymology, Lymphoma, Mantle-Cell pathology, Male, Mice, Inbred NOD, Mice, SCID, Piperidines pharmacology, Protein Kinases metabolism, RNA Polymerase II metabolism, Signal Transduction drug effects, Transcription Factors antagonists & inhibitors, Transcription Factors metabolism, Transcriptome genetics, Treatment Outcome, Mice, Adenine analogs & derivatives, Drug Resistance, Neoplasm genetics, Lymphoma, Mantle-Cell drug therapy, Lymphoma, Mantle-Cell genetics, Piperidines therapeutic use, Transcription, Genetic drug effects
- Abstract
Ibrutinib, a bruton's tyrosine kinase (BTK) inhibitor, provokes robust clinical responses in aggressive mantle cell lymphoma (MCL), yet many patients relapse with lethal Ibrutinib-resistant (IR) disease. Here, using genomic, chemical proteomic, and drug screen profiling, we report that enhancer remodeling-mediated transcriptional activation and adaptive signaling changes drive the aggressive phenotypes of IR. Accordingly, IR MCL cells are vulnerable to inhibitors of the transcriptional machinery and especially so to inhibitors of cyclin-dependent kinase 9 (CDK9), the catalytic subunit of the positive transcription elongation factor b (P-TEFb) of RNA polymerase II (RNAPII). Further, CDK9 inhibition disables reprogrammed signaling circuits and prevents the emergence of IR in MCL. Finally, and importantly, we find that a robust and facile ex vivo image-based functional drug screening platform can predict clinical therapeutic responses of IR MCL and identify vulnerabilities that can be targeted to disable the evolution of IR., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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28. TFEB links MYC signaling to epigenetic control of myeloid differentiation and acute myeloid leukemia.
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Yun S, Vincelette ND, Yu X, Watson GW, Fernandez MR, Yang C, Hitosugi T, Cheng CH, Freischel AR, Zhang L, Li W, Hou H, Schaub FX, Vedder AR, Cen L, McGraw KL, Moon J, Murphy DJ, Ballabio A, Kaufmann SH, Berglund AE, and Cleveland JL
- Subjects
- Cell Differentiation genetics, Epigenesis, Genetic, Humans, Mutation, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, Isocitrate Dehydrogenase genetics, Leukemia, Myeloid, Acute genetics, Proto-Oncogene Proteins c-myc genetics, Signal Transduction
- Abstract
MYC oncoproteins regulate transcription of genes directing cell proliferation, metabolism and tumorigenesis. A variety of alterations drive MYC expression in acute myeloid leukemia (AML) and enforced MYC expression in hematopoietic progenitors is sufficient to induce AML. Here we report that AML and myeloid progenitor cell growth and survival rely on MYC-directed suppression of Transcription Factor EB (TFEB), a master regulator of the autophagy-lysosome pathway. Notably, although originally identified as an oncogene, TFEB functions as a tumor suppressor in AML, where it provokes AML cell differentiation and death. These responses reflect TFEB control of myeloid epigenetic programs, by inducing expression of isocitrate dehydrogenase-1 (IDH1) and IDH2, resulting in global hydroxylation of 5-methycytosine. Finally, activating the TFEB-IDH1/IDH2-TET2 axis is revealed as a targetable vulnerability in AML. Thus, epigenetic control by a MYC-TFEB circuit dictates myeloid cell fate and is essential for maintenance of AML.
- Published
- 2021
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29. Comparative Genomics Reveals Distinct Immune-oncologic Pathways in African American Men with Prostate Cancer.
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Awasthi S, Berglund A, Abraham-Miranda J, Rounbehler RJ, Kensler K, Serna A, Vidal A, You S, Freeman MR, Davicioni E, Liu Y, Karnes RJ, Klein EA, Den RB, Trock BJ, Campbell JD, Einstein DJ, Gupta R, Balk S, Lal P, Park JY, Cleveland JL, Rebbeck TR, Freedland SJ, and Yamoah K
- Subjects
- Black or African American statistics & numerical data, Aged, Datasets as Topic, Epithelial-Mesenchymal Transition genetics, Epithelial-Mesenchymal Transition immunology, Follow-Up Studies, Genomics statistics & numerical data, Health Status Disparities, Humans, Male, Middle Aged, Neoplasm Recurrence, Local genetics, Neoplasm Recurrence, Local prevention & control, Prostate immunology, Prostate pathology, Prostatectomy, Prostatic Neoplasms immunology, Prostatic Neoplasms mortality, Prostatic Neoplasms therapy, Risk Assessment statistics & numerical data, Tumor Microenvironment genetics, White People genetics, White People statistics & numerical data, Black or African American genetics, Gene Expression Regulation, Neoplastic immunology, Neoplasm Recurrence, Local immunology, Prostatic Neoplasms genetics, Tumor Microenvironment immunology
- Abstract
Purpose: The role of immune-oncologic mechanisms of racial disparities in prostate cancer remains understudied. Limited research exists to evaluate the molecular underpinnings of immune differences in African American men (AAM) and European American men (EAM) prostate tumor microenvironment (TME)., Experimental Design: A total of 1,173 radiation-naïve radical prostatectomy samples with whole transcriptome data from the Decipher GRID registry were used. Transcriptomic expressions of 1,260 immune-specific genes were selected to assess immune-oncologic differences between AAM and EAM prostate tumors. Race-specific differential expression of genes was assessed using a rank test, and intergene correlational matrix and gene set enrichment was used for pathway analysis., Results: AAM prostate tumors have significant enrichment of major immune-oncologic pathways, including proinflammatory cytokines, IFNα, IFNγ, TNFα signaling, ILs, and epithelial-mesenchymal transition. AAM TME has higher total immune content score (ICS
HIGH ) compared with 0 (37.8% vs. 21.9%, P = 0.003). AAM tumors also have lower DNA damage repair and are genomically radiosensitive as compared with EAM. IFITM3 (IFN-inducible transmembrane protein 3) was one of the major proinflammatory genes overexpressed in AAM that predicted increased risk of biochemical recurrence selectively for AAM in both discovery [HRAAM = 2.30; 95% confidence interval (CI), 1.21-4.34; P = 0.01] and validation (HRAAM = 2.42; 95% CI, 1.52-3.86; P = 0.0001) but not in EAM., Conclusions: Prostate tumors of AAM manifest a unique immune repertoire and have significant enrichment of proinflammatory immune pathways that are associated with poorer outcomes. Observed immune-oncologic differences can aid in a genomically adaptive approach to treating prostate cancer in AAM., (©2020 American Association for Cancer Research.)- Published
- 2021
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30. TP53 mutations in myelodysplastic syndromes and secondary AML confer an immunosuppressive phenotype.
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Sallman DA, McLemore AF, Aldrich AL, Komrokji RS, McGraw KL, Dhawan A, Geyer S, Hou HA, Eksioglu EA, Sullivan A, Warren S, MacBeth KJ, Meggendorfer M, Haferlach T, Boettcher S, Ebert BL, Al Ali NH, Lancet JE, Cleveland JL, Padron E, and List AF
- Subjects
- Adult, Aged, Aged, 80 and over, Female, Humans, Immunosuppression Therapy, Male, MicroRNAs genetics, MicroRNAs immunology, Middle Aged, Myeloid-Derived Suppressor Cells pathology, RNA, Neoplasm genetics, RNA, Neoplasm immunology, T-Lymphocytes, Regulatory pathology, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute immunology, Leukemia, Myeloid, Acute pathology, Mutation, Myelodysplastic Syndromes genetics, Myelodysplastic Syndromes immunology, Myelodysplastic Syndromes pathology, Myeloid-Derived Suppressor Cells immunology, T-Lymphocytes, Regulatory immunology, Tumor Suppressor Protein p53 immunology
- Abstract
Somatic gene mutations are key determinants of outcome in patients with myelodysplastic syndromes (MDS) and secondary AML (sAML). In particular, patients with TP53 mutations represent a distinct molecular cohort with uniformly poor prognosis. The precise pathogenetic mechanisms underlying these inferior outcomes have not been delineated. In this study, we characterized the immunological features of the malignant clone and alterations in the immune microenvironment in patients with TP53-mutant and wild-type MDS or sAML. Notably, PDL1 expression is significantly increased in hematopoietic stem cells of patients with TP53 mutations, which is associated with MYC upregulation and marked downregulation of MYC's negative regulator miR-34a, a p53 transcription target. Notably, patients with TP53 mutations display significantly reduced numbers of bone marrow-infiltrating OX40+ cytotoxic T cells and helper T cells, as well as decreased ICOS+ and 4-1BB+ natural killer cells. Further, highly immunosuppressive regulatory T cells (Tregs) (ie, ICOShigh/PD-1-) and myeloid-derived suppressor cells (PD-1low) are expanded in cases with TP53 mutations. Finally, a higher proportion of bone marrow-infiltrating ICOShigh/PD-1- Treg cells is a highly significant independent predictor of overall survival. We conclude that the microenvironment of TP53 mutant MDS and sAML has an immune-privileged, evasive phenotype that may be a primary driver of poor outcomes and submit that immunomodulatory therapeutic strategies may offer a benefit for this molecularly defined subpopulation., (© 2020 by The American Society of Hematology.)
- Published
- 2020
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31. Ablation of polyamine catabolic enzymes provokes Purkinje cell damage, neuroinflammation, and severe ataxia.
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Zahedi K, Brooks M, Barone S, Rahmati N, Murray Stewart T, Dunworth M, Destefano-Shields C, Dasgupta N, Davidson S, Lindquist DM, Fuller CE, Smith RD, Cleveland JL, Casero RA Jr, and Soleimani M
- Subjects
- Acetyltransferases genetics, Animals, Apoptosis physiology, Ataxia genetics, Ataxia pathology, Cerebellum enzymology, Cerebellum pathology, Inflammation enzymology, Inflammation genetics, Inflammation pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Oxidoreductases Acting on CH-NH Group Donors genetics, Purkinje Cells pathology, Polyamine Oxidase, Acetyltransferases deficiency, Ataxia enzymology, Oxidoreductases Acting on CH-NH Group Donors deficiency, Purkinje Cells enzymology
- Abstract
Background: Polyamine catabolism plays a key role in maintaining intracellular polyamine pools, yet its physiological significance is largely unexplored. Here, we report that the disruption of polyamine catabolism leads to severe cerebellar damage and ataxia, demonstrating the fundamental role of polyamine catabolism in the maintenance of cerebellar function and integrity., Methods: Mice with simultaneous deletion of the two principal polyamine catabolic enzymes, spermine oxidase and spermidine/spermine N
1 -acetyltransferase (Smox/Sat1-dKO), were generated by the crossbreeding of Smox-KO (Smox-/- ) and Sat1-KO (Sat1-/- ) animals. Development and progression of tissue injury was monitored using imaging, behavioral, and molecular analyses., Results: Smox/Sat1-dKO mice are normal at birth, but develop progressive cerebellar damage and ataxia. The cerebellar injury in Smox/Sat1-dKO mice is associated with Purkinje cell loss and gliosis, leading to neuroinflammation and white matter demyelination during the latter stages of the injury. The onset of tissue damage in Smox/Sat1-dKO mice is not solely dependent on changes in polyamine levels as cerebellar injury was highly selective. RNA-seq analysis and confirmatory studies revealed clear decreases in the expression of Purkinje cell-associated proteins and significant increases in the expression of transglutaminases and markers of neurodegenerative microgliosis and astrocytosis. Further, the α-Synuclein expression, aggregation, and polyamination levels were significantly increased in the cerebellum of Smox/Sat1-dKO mice. Finally, there were clear roles of transglutaminase-2 (TGM2) in the cerebellar pathologies manifest in Smox/Sat1-dKO mice, as pharmacological inhibition of transglutaminases reduced the severity of ataxia and cerebellar injury in Smox/Sat1-dKO mice., Conclusions: These results indicate that the disruption of polyamine catabolism, via coordinated alterations in tissue polyamine levels, elevated transglutaminase activity and increased expression, polyamination, and aggregation of α-Synuclein, leads to severe cerebellar damage and ataxia. These studies indicate that polyamine catabolism is necessary to Purkinje cell survival, and for sustaining the functional integrity of the cerebellum.- Published
- 2020
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32. Cereblon harnesses Myc-dependent bioenergetics and activity of CD8+ T lymphocytes.
- Author
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Hesterberg RS, Beatty MS, Han Y, Fernandez MR, Akuffo AA, Goodheart WE, Yang C, Chang S, Colin CM, Alontaga AY, McDaniel JM, Mailloux AW, Billington JMR, Yue L, Russell S, Gillies RJ, Yun SY, Ayaz M, Lawrence NJ, Lawrence HR, Yu XZ, Fu J, Darville LN, Koomen JM, Ren X, Messina J, Jiang K, Garrett TJ, Rajadhyaksha AM, Cleveland JL, and Epling-Burnette PK
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Animals, CD8-Positive T-Lymphocytes metabolism, Cells, Cultured, Immunomodulation genetics, Melanoma, Experimental pathology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Transgenic, Adaptor Proteins, Signal Transducing physiology, CD8-Positive T-Lymphocytes physiology, Energy Metabolism genetics, Lymphocyte Activation genetics, Proto-Oncogene Proteins c-myc genetics
- Abstract
Immunomodulatory drugs, such as thalidomide and related compounds, potentiate T-cell effector functions. Cereblon (CRBN), a substrate receptor of the DDB1-cullin-RING E3 ubiquitin ligase complex, is the only molecular target for this drug class, where drug-induced, ubiquitin-dependent degradation of known "neosubstrates," such as IKAROS, AIOLOS, and CK1α, accounts for their biological activity. Far less clear is whether these CRBN E3 ligase-modulating compounds disrupt the endogenous functions of CRBN. We report that CRBN functions in a feedback loop that harnesses antigen-specific CD8+ T-cell effector responses. Specifically, Crbn deficiency in murine CD8+ T cells augments their central metabolism manifested as elevated bioenergetics, with supraphysiological levels of polyamines, secondary to enhanced glucose and amino acid transport, and with increased expression of metabolic enzymes, including the polyamine biosynthetic enzyme ornithine decarboxylase. Treatment with CRBN-modulating compounds similarly augments central metabolism of human CD8+ T cells. Notably, the metabolic control of CD8+ T cells by modulating compounds or Crbn deficiency is linked to increased and sustained expression of the master metabolic regulator MYC. Finally, Crbn-deficient T cells have augmented antigen-specific cytolytic activity vs melanoma tumor cells, ex vivo and in vivo, and drive accelerated and highly aggressive graft-versus-host disease. Therefore, CRBN functions to harness the activation of CD8+ T cells, and this phenotype can be exploited by treatment with drugs., (© 2020 by The American Society of Hematology.)
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- 2020
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33. The dual PI3Kδ/CK1ε inhibitor umbralisib exhibits unique immunomodulatory effects on CLL T cells.
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Maharaj K, Powers JJ, Achille A, Mediavilla-Varela M, Gamal W, Burger KL, Fonseca R, Jiang K, Miskin HP, Maryanski D, Monastyrskyi A, Duckett DR, Roush WR, Cleveland JL, Sahakian E, and Pinilla-Ibarz J
- Subjects
- Animals, Heterocyclic Compounds, 4 or More Rings therapeutic use, Humans, Mice, Phosphatidylinositol 3-Kinases therapeutic use, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy
- Abstract
The in-clinic phosphatidylinositol 3-kinase (PI3K) inhibitors idelalisib (CAL-101) and duvelisib (IPI-145) have demonstrated high rates of response and progression-free survival in clinical trials of B-cell malignancies, such as chronic lymphocytic leukemia (CLL). However, a high incidence of adverse events has led to frequent discontinuations, limiting the clinical development of these inhibitors. By contrast, the dual PI3Kδ/casein kinase-1-ε (CK1ε) inhibitor umbralisib (TGR-1202) also shows high rates of response in clinical trials but has an improved safety profile with fewer severe adverse events. Toxicities typical of this class of PI3K inhibitors are largely thought to be immune mediated, but they are poorly characterized. Here, we report the effects of idelalisib, duvelisib, and umbralisib on regulatory T cells (Tregs) on normal human T cells, T cells from CLL patients, and T cells in an Eμ-TCL1 adoptive transfer mouse CLL model. Ex vivo studies revealed differential effects of these PI3K inhibitors; only umbralisib treatment sustained normal and CLL-associated FoxP3+ human Tregs. Further, although all 3 inhibitors exhibit antitumor efficacy in the Eμ-TCL1 CLL model, idelalisib- or duvelisib-treated mice displayed increased immune-mediated toxicities, impaired function, and reduced numbers of Tregs, whereas Treg number and function were preserved in umbralisib-treated CLL-bearing mice. Finally, our studies demonstrate that inhibition of CK1ε can improve CLL Treg number and function. Interestingly, CK1ε inhibition mitigated impairment of CLL Tregs by PI3K inhibitors in combination treatment. These results suggest that the improved safety profile of umbralisib is due to its role as a dual PI3Kδ/CK1ε inhibitor that preserves Treg number and function., (© 2020 by The American Society of Hematology.)
- Published
- 2020
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34. Metabolic reprogramming augments potency of human pSTAT3-inhibited iTregs to suppress alloreactivity.
- Author
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Walton K, Fernandez MR, Sagatys EM, Reff J, Kim J, Lee MC, Kiluk JV, Hui JYC, McKenna D Jr, Hupp M, Forster C, Linden MA, Lawrence NJ, Lawrence HR, Pidala J, Pavletic SZ, Blazar BR, Sebti SM, Cleveland JL, Anasetti C, and Betts BC
- Subjects
- Animals, Humans, Mice, Oxidation-Reduction, Graft Rejection immunology, Graft Rejection metabolism, Graft vs Host Disease immunology, Graft vs Host Disease metabolism, STAT3 Transcription Factor physiology, T-Lymphocytes, Regulatory cytology, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism
- Abstract
Immunosuppressive donor Tregs can prevent graft-versus-host disease (GVHD) or solid-organ allograft rejection. We previously demonstrated that inhibiting STAT3 phosphorylation (pSTAT3) augments FOXP3 expression, stabilizing induced Tregs (iTregs). Here we report that human pSTAT3-inhibited iTregs prevent human skin graft rejection and xenogeneic GVHD yet spare donor antileukemia immunity. pSTAT3-inhibited iTregs express increased levels of skin-homing cutaneous lymphocyte-associated antigen, immunosuppressive GARP and PD-1, and IL-9 that supports tolerizing mast cells. Further, pSTAT3-inhibited iTregs significantly reduced alloreactive conventional T cells, Th1, and Th17 cells implicated in GVHD and tissue rejection and impaired infiltration by pathogenic Th2 cells. Mechanistically, pSTAT3 inhibition of iTregs provoked a shift in metabolism from oxidative phosphorylation (OxPhos) to glycolysis and reduced electron transport chain activity. Strikingly, cotreatment with coenzyme Q10 restored OxPhos in pSTAT3-inhibited iTregs and augmented their suppressive potency. These findings support the rationale for clinically testing the safety and efficacy of metabolically tuned, human pSTAT3-inhibited iTregs to control alloreactive T cells.
- Published
- 2020
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35. Spermine oxidase mediates Helicobacter pylori-induced gastric inflammation, DNA damage, and carcinogenic signaling.
- Author
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Sierra JC, Piazuelo MB, Luis PB, Barry DP, Allaman MM, Asim M, Sebrell TA, Finley JL, Rose KL, Hill S, Holshouser SL, Casero RA, Cleveland JL, Woster PM, Schey KL, Bimczok D, Schneider C, Gobert AP, and Wilson KT
- Subjects
- Adenocarcinoma microbiology, Animals, Cell Transformation, Neoplastic, Gastritis genetics, Gastritis microbiology, Gastritis pathology, Helicobacter Infections genetics, Helicobacter Infections pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Organoids, Oxidoreductases Acting on CH-NH Group Donors deficiency, Oxidoreductases Acting on CH-NH Group Donors genetics, Proteome, RNA, Messenger biosynthesis, Signal Transduction, Spermidine biosynthesis, Stomach Neoplasms microbiology, beta Catenin physiology, Polyamine Oxidase, Adenocarcinoma etiology, DNA Damage, Gastritis enzymology, Helicobacter Infections enzymology, Helicobacter pylori pathogenicity, Oxidoreductases Acting on CH-NH Group Donors physiology, Spermine metabolism, Stomach Neoplasms etiology
- Abstract
Helicobacter pylori infection is the main risk factor for the development of gastric cancer, the third leading cause of cancer death worldwide. H. pylori colonizes the human gastric mucosa and persists for decades. The inflammatory response is ineffective in clearing the infection, leading to disease progression that may result in gastric adenocarcinoma. We have shown that polyamines are regulators of the host response to H. pylori, and that spermine oxidase (SMOX), which metabolizes the polyamine spermine into spermidine plus H
2 O2 , is associated with increased human gastric cancer risk. We now used a molecular approach to directly address the role of SMOX, and demonstrate that Smox-deficient mice exhibit significant reductions of gastric spermidine levels and H. pylori-induced inflammation. Proteomic analysis revealed that cancer was the most significantly altered functional pathway in Smox-/- gastric organoids. Moreover, there was also less DNA damage and β-catenin activation in H. pylori-infected Smox-/- mice or gastric organoids, compared to infected wild-type animals or gastroids. The link between SMOX and β-catenin activation was confirmed in human gastric organoids that were treated with a novel SMOX inhibitor. These findings indicate that SMOX promotes H. pylori-induced carcinogenesis by causing inflammation, DNA damage, and activation of β-catenin signaling.- Published
- 2020
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36. Macrophage Metabolism of Apoptotic Cell-Derived Arginine Promotes Continual Efferocytosis and Resolution of Injury.
- Author
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Yurdagul A Jr, Subramanian M, Wang X, Crown SB, Ilkayeva OR, Darville L, Kolluru GK, Rymond CC, Gerlach BD, Zheng Z, Kuriakose G, Kevil CG, Koomen JM, Cleveland JL, Muoio DM, and Tabas I
- Subjects
- Animals, Apoptosis genetics, Arginase metabolism, ELAV-Like Protein 1 metabolism, Gene Deletion, Guanine Nucleotide Exchange Factors genetics, Guanine Nucleotide Exchange Factors metabolism, Humans, Jurkat Cells, Macrophages drug effects, Male, Mice, Inbred C57BL, Myeloid Cells drug effects, Myeloid Cells metabolism, Ornithine Decarboxylase metabolism, Phagocytosis genetics, Putrescine biosynthesis, RNA Stability drug effects, RNA Stability genetics, RNA, Messenger genetics, RNA, Messenger metabolism, rac1 GTP-Binding Protein metabolism, Apoptosis drug effects, Arginine pharmacology, Macrophages metabolism, Macrophages pathology, Phagocytosis drug effects
- Abstract
Continual efferocytic clearance of apoptotic cells (ACs) by macrophages prevents necrosis and promotes injury resolution. How continual efferocytosis is promoted is not clear. Here, we show that the process is optimized by linking the metabolism of engulfed cargo from initial efferocytic events to subsequent rounds. We found that continual efferocytosis is enhanced by the metabolism of AC-derived arginine and ornithine to putrescine by macrophage arginase 1 (Arg1) and ornithine decarboxylase (ODC). Putrescine augments HuR-mediated stabilization of the mRNA encoding the GTP-exchange factor Dbl, which activates actin-regulating Rac1 to facilitate subsequent rounds of AC internalization. Inhibition of any step along this pathway after first-AC uptake suppresses second-AC internalization, whereas putrescine addition rescues this defect. Mice lacking myeloid Arg1 or ODC have defects in efferocytosis in vivo and in atherosclerosis regression, while treatment with putrescine promotes atherosclerosis resolution. Thus, macrophage metabolism of AC-derived metabolites allows for optimal continual efferocytosis and resolution of injury., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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37. Engineered BRET-Based Biologic Light Sources Enable Spatiotemporal Control over Diverse Optogenetic Systems.
- Author
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Parag-Sharma K, O'Banion CP, Henry EC, Musicant AM, Cleveland JL, Lawrence DS, and Amelio AL
- Subjects
- HEK293 Cells, HeLa Cells, Humans, Luciferases chemistry, Transfection, Biological Products chemistry, Bioluminescence Resonance Energy Transfer Techniques methods, Green Fluorescent Proteins chemistry, Light, Luminescent Proteins chemistry, Optogenetics methods
- Abstract
Light-inducible optogenetic systems offer precise spatiotemporal control over a myriad of biologic processes. Unfortunately, current systems are inherently limited by their dependence on external light sources for their activation. Further, the utility of laser/LED-based illumination strategies are often constrained by the need for invasive surgical procedures to deliver such devices and local heat production, photobleaching and phototoxicity that compromises cell and tissue viability. To overcome these limitations, we developed a novel BRET-activated optogenetics (BEACON) system that employs biologic light to control optogenetic tools. BEACON is driven by self-illuminating bioluminescent-fluorescent proteins that generate "spectrally tuned" biologic light via bioluminescence resonance energy transfer (BRET). Notably, BEACON robustly activates a variety of commonly used optogenetic systems in a spatially restricted fashion, and at physiologically relevant time scales, to levels that are achieved by conventional laser/LED light sources.
- Published
- 2020
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38. Commercial Gene Expression Tests for Prostate Cancer Prognosis Provide Paradoxical Estimates of Race-Specific Risk.
- Author
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Creed JH, Berglund AE, Rounbehler RJ, Awasthi S, Cleveland JL, Park JY, Yamoah K, and Gerke TA
- Subjects
- Humans, Male, Neoplasm Grading, Neoplasm Staging, Predictive Value of Tests, Prostatic Neoplasms pathology, Race Factors, Black or African American genetics, Biomarkers, Tumor genetics, Prostatic Neoplasms ethnology, Prostatic Neoplasms genetics, Transcriptome, White People genetics
- Abstract
Background: Commercial gene expression signatures of prostate cancer prognosis were developed and validated in cohorts of predominantly European American men (EAM). Limited research exists on the value of such signatures in African American men (AAM), who have poor prostate cancer outcomes. We explored differences in gene expression between EAM and AAM for three commercially available panels recommended by the National Comprehensive Cancer Network for prostate cancer prognosis., Methods: A total of 232 EAM and 95 AAM patients provided radical prostatectomy specimens. Gene expression was quantified using NanoString for 60 genes spanning the Oncotype DX Prostate, Prolaris, and Decipher panels. A continuous expression-based risk score was approximated for each. Differential expression, intrapanel coexpression, and risk by race were assessed., Results: Clinical and pathologic features were similar between AAM and EAM. Differential expression by race was observed for 48% of genes measured, although the magnitudes of expression differences were small. Coexpression patterns were more strongly preserved by race group for Oncotype DX and Decipher than Prolaris. Poorer prognosis was estimated in EAM versus AAM for Oncotype DX ( P < 0.001), whereas negligible prognostic differences were predicted between AAM and EAM using Prolaris or Decipher ( P > 0.05)., Conclusions: Because of observed racial differences across three commercial gene expression panels for prostate cancer prognosis, caution is warranted when applying these panels in clinical decision-making in AAM., Impact: Differences in gene expression by race for three commercial panels for prostate cancer prognosis indicate that further study of their effectiveness in AAM with long-term follow-up is warranted., (©2019 American Association for Cancer Research.)
- Published
- 2020
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39. An Immunocompetent Mouse Model of HPV16(+) Head and Neck Squamous Cell Carcinoma.
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Carper MB, Troutman S, Wagner BL, Byrd KM, Selitsky SR, Parag-Sharma K, Henry EC, Li W, Parker JS, Montgomery SA, Cleveland JL, Williams SE, Kissil JL, Hayes DN, and Amelio AL
- Subjects
- Animals, Carcinogenesis genetics, Carcinogenesis metabolism, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases genetics, Class I Phosphatidylinositol 3-Kinases metabolism, Female, Gene Expression, Head and Neck Neoplasms genetics, Head and Neck Neoplasms immunology, Head and Neck Neoplasms metabolism, Humans, Immunocompetence, Internal Ribosome Entry Sites genetics, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Natural Killer T-Cells immunology, Natural Killer T-Cells metabolism, Oncogene Proteins, Viral genetics, Oropharyngeal Neoplasms genetics, Oropharyngeal Neoplasms immunology, Oropharyngeal Neoplasms metabolism, Papillomavirus E7 Proteins genetics, RNA Splicing genetics, RNA-Seq, Repressor Proteins genetics, Squamous Cell Carcinoma of Head and Neck genetics, Squamous Cell Carcinoma of Head and Neck immunology, Squamous Cell Carcinoma of Head and Neck metabolism, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism, Disease Models, Animal, Head and Neck Neoplasms virology, Oncogene Proteins, Viral metabolism, Oropharyngeal Neoplasms virology, Papillomavirus E7 Proteins metabolism, Repressor Proteins metabolism, Squamous Cell Carcinoma of Head and Neck virology
- Abstract
The incidence of human papilloma virus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) is increasing and implicated in more than 60% of all oropharyngeal carcinomas (OPSCCs). Although whole-genome, transcriptome, and proteome analyses have identified altered signaling pathways in HPV-induced HNSCCs, additional tools are needed to investigate the unique pathobiology of OPSCC. Herein, bioinformatics analyses of human HPV(+) HNSCCs revealed that all tumors express full-length E6 and identified molecular subtypes based on relative E6 and E7 expression levels. To recapitulate the levels, stoichiometric ratios, and anatomic location of E6/E7 expression, we generated a genetically engineered mouse model whereby balanced expression of E6/E7 is directed to the oropharyngeal epithelium. The addition of a mutant PIK3CA
E545K allele leads to the rapid development of pre-malignant lesions marked by immune cell accumulation, and a subset of these lesions progress to OPSCC. This mouse provides a faithful immunocompetent model for testing treatments and investigating mechanisms of immunosuppression., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2019
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40. Betaglycan drives the mesenchymal stromal cell osteogenic program and prostate cancer-induced osteogenesis.
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Cook LM, Frieling JS, Nerlakanti N, McGuire JJ, Stewart PA, Burger KL, Cleveland JL, and Lynch CC
- Subjects
- Humans, Male, Proteoglycans genetics, Receptors, Transforming Growth Factor beta genetics, Wnt-5a Protein metabolism, Mesenchymal Stem Cells pathology, Osteoblasts pathology, Osteogenesis, Prostatic Neoplasms pathology, Proteoglycans metabolism, Receptors, Transforming Growth Factor beta metabolism
- Abstract
Bone metastatic prostate cancer provokes extensive osteogenesis by driving the recruitment and osteoblastic differentiation of mesenchymal stromal cells (MSCs). The resulting lesions greatly contribute to patient morbidity and mortality, underscoring the need for defining how prostate metastases subvert the MSC-osteoblast differentiation program. To gain insights into this process we profiled the effects of co-culture of primary MSCs with validated bone metastatic prostate cancer cell line models. These analyses revealed a cast of shared differentially induced genes in MSC, including betaglycan, a co-receptor for TGFβ. Betaglycan has not been studied in the context of bone metastatic disease previously. Here we report that loss of betaglycan in MSC is sufficient to augment TGFβ signaling, proliferation and migration, and completely blocks the MSC-osteoblast differentiation program. Further, betaglycan was revealed as necessary for prostate cancer-induced osteogenesis in vivo. Mechanistically, gene expression analysis revealed betaglycan controls the expression of a large repertoire of genes in MSCs, and that betaglycan loss provokes >60-fold increase in the expression of Wnt5a that plays important roles in stemness. In accord with the increased Wnt5a levels, there was a marked induction of canonical Wnt signaling in betaglycan ablated MSCs, and the addition of recombinant Wnt5a to MSCs was sufficient to impair osteogenic differentiation. Finally, the addition of Wnt5a neutralizing antibody was sufficient to induce the expression of osteogenic genes in betaglycan-ablated MSCs. Collectively, these findings suggest a betaglycan-Wnt5a circuit represents an attractive vulnerability to ameliorate prostate cancer-induced osteogenesis.
- Published
- 2019
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41. Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy.
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Renner K, Bruss C, Schnell A, Koehl G, Becker HM, Fante M, Menevse AN, Kauer N, Blazquez R, Hacker L, Decking SM, Bohn T, Faerber S, Evert K, Aigle L, Amslinger S, Landa M, Krijgsman O, Rozeman EA, Brummer C, Siska PJ, Singer K, Pektor S, Miederer M, Peter K, Gottfried E, Herr W, Marchiq I, Pouyssegur J, Roush WR, Ong S, Warren S, Pukrop T, Beckhove P, Lang SA, Bopp T, Blank CU, Cleveland JL, Oefner PJ, Dettmer K, Selby M, and Kreutz M
- Subjects
- Animals, Anti-Inflammatory Agents, Non-Steroidal pharmacology, Cell Line, Tumor, Cell Proliferation drug effects, Cell Proliferation physiology, Glucose metabolism, Glycolysis drug effects, Humans, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, T-Lymphocytes drug effects, T-Lymphocytes metabolism, Xenopus laevis, Glycolysis physiology, T-Lymphocytes physiology
- Abstract
Tumor-derived lactic acid inhibits T and natural killer (NK) cell function and, thereby, tumor immunosurveillance. Here, we report that melanoma patients with high expression of glycolysis-related genes show a worse progression free survival upon anti-PD1 treatment. The non-steroidal anti-inflammatory drug (NSAID) diclofenac lowers lactate secretion of tumor cells and improves anti-PD1-induced T cell killing in vitro. Surprisingly, diclofenac, but not other NSAIDs, turns out to be a potent inhibitor of the lactate transporters monocarboxylate transporter 1 and 4 and diminishes lactate efflux. Notably, T cell activation, viability, and effector functions are preserved under diclofenac treatment and in a low glucose environment in vitro. Diclofenac, but not aspirin, delays tumor growth and improves the efficacy of checkpoint therapy in vivo. Moreover, genetic suppression of glycolysis in tumor cells strongly improves checkpoint therapy. These findings support the rationale for targeting glycolysis in patients with high glycolytic tumors together with checkpoint inhibitors in clinical trials., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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42. Prognostic significance of MYC oncoprotein expression on survival outcome in patients with acute myeloid leukemia with myelodysplasia related changes (AML-MRC).
- Author
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Yun S, Sharma R, Chan O, Vincelette ND, Sallman DA, Sweet K, Padron E, Komrokji R, Lancet JE, Abraham I, Moscinski LC, Cleveland JL, List AF, and Zhang L
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Biopsy, Chromosome Aberrations, Female, Humans, Immunohistochemistry, Leukemia, Myeloid, Acute pathology, Leukemia, Myeloid, Acute therapy, Male, Middle Aged, Mutation, Myelodysplastic Syndromes pathology, Myelodysplastic Syndromes therapy, Prognosis, Proportional Hazards Models, Gene Expression Regulation, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute mortality, Myelodysplastic Syndromes genetics, Myelodysplastic Syndromes mortality, Proto-Oncogene Proteins c-myc genetics
- Abstract
MYC is an oncoprotein that coordinates the expression of genes involved in metabolism, cell differentiation and survival in various types of malignancies. However, the underlying oncogenic mechanisms and the clinical significance of MYC expression in the acute myeloid leukemia with myelodysplasia related changes (AML-MRC) remain to be answered. A total of 135 patients were retrospectively identified using Total Cancer Care (TCC) Moffitt Cancer Center (MCC) databases. Diagnosis of AML-MRC was based on the 2016 WHO classification utilizing bone marrow (BM) evaluation. MYC protein expression level was assessed by immunohistochemistry (IHC) staining using paraffin-embedded BM trephine biopsy samples obtained at the time of diagnosis or relapse. Concurrent somatic mutations were assessed using targeted next generation sequencing that include 54 genes. A total of 38% (n = 51) and 62% (n = 84) patients had high and low MYC expression, respectively. The most common somatic mutation in our cohort was TP53 followed by DNMT3A, and ASXL1. The median OS was significantly longer in low MYC patients (median OS 42.3 vs. 17.05 months, p = 0.0109). Multivariate analysis including MYC expression level, transplantation status, gender and age demonstrated high MYC expression (HR 1.77, 95% CI 1.004-3.104, p = 0.045) to be an independent poor prognostic factor. Further studies are needed to identify the underlying mechanism of MYC driven oncogenesis in AML-MRC. Additionally, the prognostic impact of MYC on the AML survival in a larger cohort that include diverse somatic mutations and chromosomal abnormalities requires further investigation., (Copyright © 2019. Published by Elsevier Ltd.)
- Published
- 2019
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43. S100A9-induced overexpression of PD-1/PD-L1 contributes to ineffective hematopoiesis in myelodysplastic syndromes.
- Author
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Cheng P, Eksioglu EA, Chen X, Kandell W, Le Trinh T, Cen L, Qi J, Sallman DA, Zhang Y, Tu N, Adams WA, Zhang C, Liu J, Cleveland JL, List AF, and Wei S
- Subjects
- Animals, Apoptosis, B7-H1 Antigen analysis, B7-H1 Antigen antagonists & inhibitors, Humans, Mice, Mice, Inbred C57BL, Myelodysplastic Syndromes drug therapy, Programmed Cell Death 1 Receptor analysis, Programmed Cell Death 1 Receptor antagonists & inhibitors, Proto-Oncogene Proteins c-myc physiology, B7-H1 Antigen physiology, Calgranulin B physiology, Hematopoiesis, Myelodysplastic Syndromes etiology, Programmed Cell Death 1 Receptor physiology
- Abstract
Myelodysplastic syndromes (MDS) are characterized by dysplastic and ineffective hematopoiesis that can result from aberrant expansion and activation of myeloid-derived suppressor cells (MDSCs) within the bone marrow (BM) niche. MDSCs produce S100A9, which mediates premature death of hematopoietic stem and progenitor cells (HSPCs). The PD-1/PD-L1 immune checkpoint impairs immune responses by inducing T-cell exhaustion and apoptosis, but its role in MDS is uncharacterized. Here we report an increased expression of PD-1 on HSPCs and PD-L1 on MDSCs in MDS versus healthy donors, and that this checkpoint is also activated in S100A9 transgenic (S100A9Tg) mice, and by treatment of BM mononuclear cells (BM-MNC) with S100A9. Further, MDS BM-MNC treated with recombinant PD-L1 underwent cell death, suggesting that the PD-1/PD-L1 interaction contributes to HSPC death in MDS. In accordance with this notion, PD-1/PD-L1 blockade restores effective hematopoiesis and improves colony-forming capacity in BM-MNC from MDS patients. Similar findings were observed in aged S100A9Tg mice. Finally, we demonstrate that c-Myc is required for S100A9-induced upregulation of PD-1/PD-L1, and that treatment of MDS HSPCs with anti-PD-1 antibody suppresses the expression of Myc target genes and increases the expression of hematopoietic pathway genes. We conclude anti-PD-1/anti-PD-L1 blocking strategies offer therapeutic promise in MDS in restoring effective hematopoiesis.
- Published
- 2019
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44. African American Specific Gene Panel Predictive of Poor Prostate Cancer Outcome.
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Echevarria MI, Awasthi S, Cheng CH, Berglund AE, Rounbehler RJ, Gerke TA, Takhar M, Davicioni E, Klein EA, Freedland SJ, Ross AE, Schaeffer EM, Den RB, Cleveland JL, Park JY, Rayford W, and Yamoah K
- Subjects
- Aged, Biomarkers, Tumor genetics, Cohort Studies, Gene Expression Regulation, Humans, Male, Middle Aged, Neoplasm Recurrence, Local genetics, Prognosis, Proto-Oncogene Proteins c-ets genetics, Black or African American genetics, Prostatic Neoplasms genetics
- Abstract
Purpose: Most prostate cancer in African American men lacks the ETS (E26 transforming specific) family fusion event (ETS-). We aimed to establish clinically relevant biomarkers in African American men by studying ETS dependent gene expression patterns to identified race specific genes predictive of outcomes., Materials and Methods: Two multicenter cohorts of a total of 1,427 men were used for the discovery and validation (635 and 792 men, respectively) of race specific predictive biomarkers. We used false discovery rate adjusted q values to identify race and ETS dependent genes which were differentially expressed in African American men who experienced biochemical recurrence within 5 years. Principal component modeling along with survival analysis was done to assess the accuracy of the gene panel in predicting recurrence., Results: We identified 3,047 genes which were differentially expressed based on ETS status. Of these genes 362 were differentially expressed in a race specific manner (false discovery rate 0.025 or less). A total of 81 genes were race specific and over expressed in African American men who experienced biochemical recurrence. The final gene panel included APOD, BCL6, EMP1, MYADM, SRGN and TIMP3. These genes were associated with 5-year biochemical recurrence (HR 1.97, 95% CI 1.27-3.06, p = 0.002) and they improved the predictive accuracy of clinicopathological variables only in African American men (60-month time dependent AUC 0.72)., Conclusions: In an effort to elucidate biological features associated with prostate cancer aggressiveness in African American men we identified ETS dependent biomarkers predicting early onset biochemical recurrence only in African American men. Thus, these ETS dependent biomarkers representing ideal candidates for biomarkers of aggressive disease in this patient population.
- Published
- 2019
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45. BCL2 Amplicon Loss and Transcriptional Remodeling Drives ABT-199 Resistance in B Cell Lymphoma Models.
- Author
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Zhao X, Ren Y, Lawlor M, Shah BD, Park PMC, Lwin T, Wang X, Liu K, Wang M, Gao J, Li T, Xu M, Silva AS, Lee K, Zhang T, Koomen JM, Jiang H, Sudalagunta PR, Meads MB, Cheng F, Bi C, Fu K, Fan H, Dalton WS, Moscinski LC, Shain KH, Sotomayor EM, Wang GG, Gray NS, Cleveland JL, Qi J, and Tao J
- Abstract
Drug-tolerant "persister" tumor cells underlie emergence of drug-resistant clones and contribute to relapse and disease progression. Here we report that resistance to the BCL-2 targeting drug ABT-199 in models of mantle cell lymphoma and double-hit lymphoma evolves from outgrowth of persister clones displaying loss of 18q21 amplicons that harbor BCL2. Further, persister status is generated via adaptive super-enhancer remodeling that reprograms transcription and offers opportunities for overcoming ABT-199 resistance. Notably, pharmacoproteomic and pharmacogenomic screens revealed that persisters are vulnerable to inhibition of the transcriptional machinery and especially to inhibition of cyclin-dependent kinase 7 (CDK7), which is essential for the transcriptional reprogramming that drives and sustains ABT-199 resistance. Thus, transcription-targeting agents offer new approaches to disable drug resistance in B-cell lymphomas., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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46. Distinct transcriptional repertoire of the androgen receptor in ETS fusion-negative prostate cancer.
- Author
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Berglund AE, Rounbehler RJ, Gerke T, Awasthi S, Cheng CH, Takhar M, Davicioni E, Alshalalfa M, Erho N, Klein EA, Freedland SJ, Ross AE, Schaeffer EM, Trock BJ, Den RB, Cleveland JL, Park JY, Dhillon J, and Yamoah K
- Subjects
- Cell Line, Tumor, Computational Biology methods, Gene Expression Profiling, Gene Ontology, Humans, Male, Neoplasm Grading, Neoplasm Staging, Prostatic Neoplasms pathology, Gene Expression Regulation, Neoplastic, Oncogene Proteins, Fusion genetics, Prostatic Neoplasms genetics, Prostatic Neoplasms metabolism, Proto-Oncogene Proteins c-ets genetics, Receptors, Androgen metabolism, Transcriptome
- Abstract
Background: Prostate cancer (PCa) tumors harboring translocations of ETS family genes with the androgen responsive TMPRSS2 gene (ETS+ tumors) provide a robust biomarker for detecting PCa in approximately 70% of patients. ETS+ PCa express high levels of the androgen receptor (AR), yet PCa tumors lacking ETS fusions (ETS-) also express AR and demonstrate androgen-regulated growth. In this study, we evaluate the differences in the AR-regulated transcriptomes between ETS+ and ETS- PCa tumors., Methods: 10,608 patient tumors from three independent PCa datasets classified as ETS+ (samples overexpressing ERG or other ETS family members) or ETS- (all other PCa) were analyzed for differential gene expression using false-discovery-rate adjusted methods and gene-set enrichment analysis (GSEA)., Results: Based on the expression of AR-dependent genes and an unsupervised Principal Component Analysis (PCA) model, AR-regulated gene expression alone was able to separate PCa samples into groups based on ETS status in all PCa databases. ETS status distinguished several differentially expressed genes in both TCGA (6.9%) and GRID (6.6%) databases, with 413 genes overlapping in both databases. Importantly, GSEA showed enrichment of distinct androgen-responsive genes in both ETS- and ETS+ tumors, and AR ChIP-seq data identified 131 direct AR-target genes that are regulated in an ETS-specific fashion. Notably, dysregulation of ETS-dependent AR-target genes within the metabolic and non-canonical WNT pathways was associated with clinical outcomes., Conclusions: ETS status influences the transcriptional repertoire of the AR, and ETS- PCa tumors appear to rely on distinctly different AR-dependent transcriptional programs to drive and sustain tumorigenesis.
- Published
- 2019
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47. PLK1 stabilizes a MYC-dependent kinase network in aggressive B cell lymphomas.
- Author
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Ren Y, Bi C, Zhao X, Lwin T, Wang C, Yuan J, Silva AS, Shah BD, Fang B, Li T, Koomen JM, Jiang H, Chavez JC, Pham LV, Sudalagunta PR, Wan L, Wang X, Dalton WS, Moscinski LC, Shain KH, Vose J, Cleveland JL, Sotomayor EM, Fu K, and Tao J
- Subjects
- Animals, Cell Cycle Proteins genetics, Cell Line, Tumor, Female, Humans, Lymphoma, Large B-Cell, Diffuse genetics, Lymphoma, Large B-Cell, Diffuse pathology, Male, Mice, Mice, Inbred NOD, Mice, SCID, Myeloid Cell Leukemia Sequence 1 Protein genetics, Myeloid Cell Leukemia Sequence 1 Protein metabolism, Protein Serine-Threonine Kinases genetics, Protein Stability, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins c-myc genetics, Polo-Like Kinase 1, Cell Cycle Proteins metabolism, Lymphoma, Large B-Cell, Diffuse metabolism, Protein Serine-Threonine Kinases metabolism, Proteolysis, Proto-Oncogene Proteins metabolism, Proto-Oncogene Proteins c-myc metabolism, Signal Transduction
- Abstract
Concordant activation of MYC and BCL-2 oncoproteins in double-hit lymphoma (DHL) results in aggressive disease that is refractory to treatment. By integrating activity-based proteomic profiling and drug screens, polo-like kinase-1 (PLK1) was identified as an essential regulator of the MYC-dependent kinome in DHL. Notably, PLK1 was expressed at high levels in DHL, correlated with MYC expression, and connoted poor outcome. Further, PLK1 signaling augmented MYC protein stability, and in turn, MYC directly induced PLK1 transcription, establishing a feed-forward MYC-PLK1 circuit in DHL. Finally, inhibition of PLK1 triggered degradation of MYC and of the antiapoptotic protein MCL-1, and PLK1 inhibitors showed synergy with BCL-2 antagonists in blocking DHL cell growth, survival, and tumorigenicity, supporting clinical targeting of PLK1 in DHL.
- Published
- 2018
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48. Tristetraprolin Is a Prognostic Biomarker for Poor Outcomes among Patients with Low-Grade Prostate Cancer.
- Author
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Rounbehler RJ, Berglund AE, Gerke T, Takhar MM, Awasthi S, Li W, Davicioni E, Erho NG, Ross AE, Schaeffer EM, Klein EA, Karnes RJ, Jenkins RB, Cleveland JL, Park JY, and Yamoah K
- Subjects
- Humans, Male, Prostatic Neoplasms pathology, Risk Factors, Treatment Outcome, Biomarkers, Tumor metabolism, Prostatic Neoplasms genetics, Tristetraprolin genetics
- Abstract
Background: We studied the utility of the tumor suppressor Tristetraprolin (TTP, ZFP36) as a clinically relevant biomarker of aggressive disease in prostate cancer patients after radical prostatectomy (RP). Methods: TTP RNA expression was measured in an RP cohort of patients treated at Moffitt Cancer Center (MCC) and obtained from six publically available RP datasets with biochemical recurrence (BCR; total n = 1,394) and/or metastatic outcome data (total n = 1,222). TTP protein expression was measured by immunohistochemistry in a tissue microarray of 153 MCC RP samples. The time to BCR or metastasis based on TTP RNA or protein levels was calculated using the Kaplan-Meier analysis. Univariable and multivariable Cox proportional hazard models were performed on multiple cohorts to evaluate if TTP is a clinically relevant biomarker and to assess if TTP improves upon the Cancer of the Prostate Risk Assessment postsurgical (CAPRA-S) score for predicting clinical outcomes. Results: In all of the RP patient cohorts, prostate cancer with low TTP RNA or protein levels had decreased time to BCR or metastasis versus TTP-high tumors. Further, the decreased time to BCR in TTP-low prostate cancer was more pronounced in low-grade tumors. Finally, pooled survival analysis suggests that TTP RNA expression provides independent information beyond CAPRA-S to predict BCR. Conclusions: TTP is a promising prostate cancer biomarker for predicting which RP patients will have poor outcomes, especially for low-grade prostate cancer patients. Impact: This study suggests that TTP RNA expression can be used to enhance the accuracy of CAPRA-S to predict outcomes in patients treated with RP. Cancer Epidemiol Biomarkers Prev; 27(11); 1376-83. ©2018 AACR., (©2018 American Association for Cancer Research.)
- Published
- 2018
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49. ATF4-amino acid circuits: a recipe for resistance in melanoma.
- Author
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Fernandez MR and Cleveland JL
- Subjects
- Activating Transcription Factor 4, Amino Acids, Cell Survival, Humans, Melanoma, Signal Transduction
- Published
- 2018
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50. P450-Catalyzed Tailoring Steps in Leinamycin Biosynthesis Featuring Regio- and Stereoselective Hydroxylations and Substrate Promiscuities.
- Author
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Kwong T, Ma M, Pan G, Hindra, Yang D, Yang C, Lohman JR, Rudolf JD, Cleveland JL, and Shen B
- Subjects
- Antibiotics, Antineoplastic chemistry, Antibiotics, Antineoplastic toxicity, Biosynthetic Pathways, Cell Line, Tumor, Cytochrome P-450 Enzyme System genetics, Escherichia coli genetics, Gene Silencing, Humans, Hydroxylation, Lactams chemistry, Lactams toxicity, Lactams, Macrocyclic chemistry, Lactams, Macrocyclic toxicity, Macrolides chemistry, Macrolides toxicity, Molecular Structure, Multigene Family, Stereoisomerism, Streptomyces genetics, Structure-Activity Relationship, Thiazoles chemistry, Thiazoles toxicity, Thiones chemistry, Thiones toxicity, Antibiotics, Antineoplastic biosynthesis, Cytochrome P-450 Enzyme System metabolism, Lactams metabolism, Lactams, Macrocyclic metabolism, Macrolides metabolism, Thiazoles metabolism, Thiones metabolism
- Abstract
Leinamycin (LNM) is a potent antitumor antibiotic produced by Streptomyces atroolivaceus S-140. Both in vivo and in vitro characterization of the LNM biosynthetic machinery have established the formation of the 18-membered macrolactam backbone and the C-3 alkyl branch; the nascent product, LNM E1, of the hybrid nonribosomal peptide synthetase (NRPS)-acyltransferase (AT)-less type I polyketide synthase (PKS); and the generation of the thiol moiety at C-3 of LNM E1. However, the tailoring steps converting LNM E1 to LNM are still unknown. Based on gene inactivation and chemical investigation of three mutant strains, we investigated the tailoring steps catalyzed by two cytochromes P450 (P450s), LnmA and LnmZ, in LNM biosynthesis. Our studies revealed that (i) LnmA and LnmZ regio- and stereoselectively hydroxylate the C-8 and C-4' positions, respectively, on the scaffold of LNM; (ii) both LnmA and LnmZ exhibit substrate promiscuity, resulting in multiple LNM analogs from several shunt pathways; and (iii) the C-8 and C-4' hydroxyl groups play important roles in the cytotoxicity of LNM analogs against different cancer cell lines, shedding light on the structure-activity relationships of the LNM scaffold and the LNM-type natural products in general. These studies set the stage for future biosynthetic pathway engineering and combinatorial biosynthesis of the LNM family of natural products for structure diversity and drug discovery.
- Published
- 2018
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