Search

Your search keyword '"David W. Speicher"' showing total 429 results
Start Over Author "David W. Speicher"
429 results on '"David W. Speicher"'

1. HRS phosphorylation drives immunosuppressive exosome secretion and restricts CD8+ T-cell infiltration into tumors

Author

Lei Guan, Bin Wu, Ting Li, Lynn A. Beer, Gaurav Sharma, Mingyue Li, Chin Nien Lee, Shujing Liu, Changsong Yang, Lili Huang, Dennie T. Frederick, Genevieve M. Boland, Guangcan Shao, Tatyana M. Svitkina, Kathy Q. Cai, Fangping Chen, Meng-Qiu Dong, Gordon B. Mills, Lynn M. Schuchter, Giorgos C. Karakousis, Tara C. Mitchell, Keith T. Flaherty, David W. Speicher, Youhai H. Chen, Meenhard Herlyn, Ravi K. Amaravadi, Xiaowei Xu, and Wei Guo

Subjects
Science
Abstract

Lack of CD8+ T-cell infiltration into solid tumors is associated with poor responsiveness to immune checkpoint therapy (ICT). Here, the authors show that blocking the phosphorylation of HRS to reduce the induction of immunosuppressive exosomes promotes CD8+ T-cell infiltration into tumors and enhances the efficacy of ICT in mouse melanoma models.

Published
2022
Full Text
View/download PDF

2. Lysosomal lipid peroxidation regulates tumor immunity

Author

Monika Bhardwaj, Jennifer J. Lee, Amanda M. Versace, Sandra L. Harper, Aaron R. Goldman, Mary Ann S. Crissey, Vaibhav Jain, Mahendra Pal Singh, Megane Vernon, Andrew E. Aplin, Seokwoo Lee, Masao Morita, Jeffrey D. Winkler, Qin Liu, David W. Speicher, and Ravi K. Amaravadi

Subjects
Oncology, Medicine
Abstract

Lysosomal inhibition elicited by palmitoyl-protein thioesterase 1 (PPT1) inhibitors such as DC661 can produce cell death, but the mechanism for this is not completely understood. Programmed cell death pathways (autophagy, apoptosis, necroptosis, ferroptosis, and pyroptosis) were not required to achieve the cytotoxic effect of DC661. Inhibition of cathepsins, or iron or calcium chelation, did not rescue DC661-induced cytotoxicity. PPT1 inhibition induced lysosomal lipid peroxidation (LLP), which led to lysosomal membrane permeabilization and cell death that could be reversed by the antioxidant N-acetylcysteine (NAC) but not by other lipid peroxidation antioxidants. The lysosomal cysteine transporter MFSD12 was required for intralysosomal transport of NAC and rescue of LLP. PPT1 inhibition produced cell-intrinsic immunogenicity with surface expression of calreticulin that could only be reversed with NAC. DC661-treated cells primed naive T cells and enhanced T cell–mediated toxicity. Mice vaccinated with DC661-treated cells engendered adaptive immunity and tumor rejection in “immune hot” tumors but not in “immune cold” tumors. These findings demonstrate that LLP drives lysosomal cell death, a unique immunogenic form of cell death, pointing the way to rational combinations of immunotherapy and lysosomal inhibition that can be tested in clinical trials.

Published
2023
Full Text
View/download PDF

3. Proteome-defined changes in cellular pathways for decidua and trophoblast tissues associated with location and viability of early-stage pregnancy

Author

Lynn A. Beer, Suneeta Senapati, Mary D. Sammel, Kurt T. Barnhart, Courtney A. Schreiber, and David W. Speicher

Subjects
Ectopic pregnancy, Early pregnancy loss, Decidua, Trophoblast, Maternal–fetal interface, Proteomics, Gynecology and obstetrics, RG1-991, Reproduction, QH471-489
Abstract

Abstract Background In early pregnancy, differentiating between a normal intrauterine pregnancy (IUP) and abnormal gestations including early pregnancy loss (EPL) or ectopic pregnancy (EP) is a major clinical challenge when ultrasound is not yet diagnostic. Clinical treatments for these outcomes are drastically different making early, accurate diagnosis imperative. Hence, a greater understanding of the biological mechanisms involved in these early pregnancy complications could lead to new molecular diagnostics. Methods Trophoblast and endometrial tissue was collected from consenting women having an IUP (n = 4), EPL (n = 4), or EP (n = 2). Samples were analyzed by LC–MS/MS followed by a label-free proteomics analysis in an exploratory study. For each tissue type, pairwise comparisons of different pregnancy outcomes (EPL vs. IUP and EP vs. IUP) were performed, and protein changes having a fold change ≥ 3 and a Student’s t-test p-value ≤ 0.05 were defined as significant. Pathway and network classification tools were used to group significantly changing proteins based on their functional similarities. Results A total of 4792 and 4757 proteins were identified in decidua and trophoblast proteomes. For decidua, 125 protein levels (2.6% of the proteome) were significantly different between EP and IUP, whereas EPL and IUP decidua were more similar with only 68 (1.4%) differences. For trophoblasts, there were 66 (1.4%) differences between EPL and IUP. However, the largest group of 344 differences (7.2%) was observed between EP and IUP trophoblasts. In both tissues, proteins associated with ECM remodeling, cell adhesion and metabolic pathways showed decreases in EP specimens compared with IUP and EPL. In trophoblasts, EP showed elevation of inflammatory and immune response pathways. Conclusions Overall, differences between an EP and IUP are greater than the changes observed when comparing ongoing IUP and nonviable intrauterine pregnancies (EPL) in both decidua and trophoblast proteomes. Furthermore, differences between EP and IUP were much higher in the trophoblast than in the decidua. This observation is true for the total number of protein changes as well as the extent of changes in upstream regulators and related pathways. This suggests that biomarkers and mechanisms of trophoblast function may be the best predictors of early pregnancy location and viability.

Published
2022
Full Text
View/download PDF

5. Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular metastasis and enhances cell death induced by DNA damaging chemotherapy

Author

Xuanmao Jiao, Min Wang, Zhao Zhang, Zhiping Li, Dong Ni, Anthony W. Ashton, Hsin-Yao Tang, David W. Speicher, and Richard G. Pestell

Subjects
Leronlimab, CCR5, Metastasis, Breast cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Abstract Background Triple-negative breast cancer (BCa) (TNBC) is a deadly form of human BCa with limited treatment options and poor prognosis. In our prior analysis of over 2200 breast cancer samples, the G protein-coupled receptor CCR5 was expressed in > 95% of TNBC samples. A humanized monoclonal antibody to CCR5 (leronlimab), used in the treatment of HIV-infected patients, has shown minimal side effects in large patient populations. Methods A humanized monoclonal antibody to CCR5, leronlimab, was used for the first time in tissue culture and in mice to determine binding characteristics to human breast cancer cells, intracellular signaling, and impact on (i) metastasis prevention and (ii) impact on established metastasis. Results Herein, leronlimab was shown to bind CCR5 in multiple breast cancer cell lines. Binding of leronlimab to CCR5 reduced ligand-induced Ca+ 2 signaling, invasion of TNBC into Matrigel, and transwell migration. Leronlimab enhanced the BCa cell killing of the BCa chemotherapy reagent, doxorubicin. In xenografts conducted with Nu/Nu mice, leronlimab reduced lung metastasis of the TNBC cell line, MB-MDA-231, by > 98% at 6 weeks. Treatment with leronlimab reduced the metastatic tumor burden of established TNBC lung metastasis. Conclusions The safety profile of leronlimab, together with strong preclinical evidence to both prevent and reduce established breast cancer metastasis herein, suggests studies of clinical efficacy may be warranted.

Published
2021
Full Text
View/download PDF

6. Topoisomerase 1 cleavage complex enables pattern recognition and inflammation during senescence

Author

Bo Zhao, Pingyu Liu, Takeshi Fukumoto, Timothy Nacarelli, Nail Fatkhutdinov, Shuai Wu, Jianhuang Lin, Katherine M. Aird, Hsin-Yao Tang, Qin Liu, David W. Speicher, and Rugang Zhang

Subjects
Science
Abstract

Here, the authors show that the topoisomerase 1-DNA covalent cleavage complex plays a critical role in mediating cytoplasmic chromatin fragments recognition by cyclic GMP-AMP synthase during senescence. The proposed axis is crucial to promote the inflammatory senescence-associated secretory phenotype and to enable the response to immune checkpoint blockade.

Published
2020
Full Text
View/download PDF

7. Protein Posttranslational Signatures Identified in COVID-19 Patient Plasma

Author

Pavan Vedula, Hsin-Yao Tang, David W. Speicher, Anna Kashina, and The UPenn COVID Processing Unit

Subjects
COVID-19, proteomics, peptidomics, posttranslational modifications, arginylation, Biology (General), QH301-705.5
Abstract

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a highly contagious virus of the coronavirus family that causes coronavirus disease-19 (COVID-19) in humans and a number of animal species. COVID-19 has rapidly propagated in the world in the past 2 years, causing a global pandemic. Here, we performed proteomic analysis of plasma samples from COVID-19 patients compared to healthy control donors in an exploratory study to gain insights into protein-level changes in the patients caused by SARS-CoV-2 infection and to identify potential proteomic and posttranslational signatures of this disease. Our results suggest a global change in protein processing and regulation that occurs in response to SARS-CoV-2, and the existence of a posttranslational COVID-19 signature that includes an elevation in threonine phosphorylation, a change in glycosylation, and a decrease in arginylation, an emerging posttranslational modification not previously implicated in infectious disease. This study provides a resource for COVID-19 researchers and, longer term, and will inform our understanding of this disease and its treatment.

Published
2022
Full Text
View/download PDF

8. PPT1 inhibition enhances the antitumor activity of anti–PD-1 antibody in melanoma

Author

Gaurav Sharma, Rani Ojha, Estela Noguera-Ortega, Vito W. Rebecca, John Attanasio, Shujing Liu, Shengfu Piao, Jennifer J. Lee, Michael C. Nicastri, Sandra L. Harper, Amruta Ronghe, Vaibhav Jain, Jeffrey D. Winkler, David W. Speicher, Jerome Mastio, Phyllis A. Gimotty, Xiaowei Xu, E. John Wherry, Dmitry I. Gabrilovich, and Ravi K. Amaravadi

Subjects
Oncology, Therapeutics, Medicine
Abstract

New strategies are needed to enhance the efficacy of anti–programmed cell death protein antibody (anti–PD-1 Ab) in cancer. Here, we report that inhibiting palmitoyl-protein thioesterase 1 (PPT1), a target of chloroquine derivatives like hydroxychloroquine (HCQ), enhances the antitumor efficacy of anti–PD-1 Ab in melanoma. The combination resulted in tumor growth impairment and improved survival in mouse models. Genetic suppression of core autophagy genes, but not Ppt1, in cancer cells reduced priming and cytotoxic capacity of primed T cells. Exposure of antigen-primed T cells to macrophage-conditioned medium derived from macrophages treated with PPT1 inhibitors enhanced melanoma-specific killing. Genetic or chemical Ppt1 inhibition resulted in M2 to M1 phenotype switching in macrophages. The combination was associated with a reduction in myeloid-derived suppressor cells in the tumor. Ppt1 inhibition by HCQ, or DC661, induced cyclic GMP-AMP synthase/stimulator of interferon genes/TANK binding kinase 1 pathway activation and the secretion of interferon-β in macrophages, the latter being a key component for augmented T cell–mediated cytotoxicity. Genetic Ppt1 inhibition produced similar findings. These data provide the rationale for this combination in melanoma clinical trials and further investigation in other cancers.

Published
2020
Full Text
View/download PDF

9. SWI/SNF catalytic subunits’ switch drives resistance to EZH2 inhibitors in ARID1A-mutated cells

Author

Shuai Wu, Nail Fatkhutdinov, Takeshi Fukumoto, Benjamin G. Bitler, Pyoung Hwa Park, Andrew V. Kossenkov, Marco Trizzino, Hsin-Yao Tang, Lin Zhang, Alessandro Gardini, David W. Speicher, and Rugang Zhang

Subjects
Science
Abstract

The mechanism of resistance to EZH2 inhibitors in cancers with inactivating SWI/SNF mutations is unknown. Here, the authors demonstrate that the switch of the SWI/SNF catalytic subunits from SMARCA4 to SMARCA2 drives resistance to EZH2 inhibitors in ARID1A-mutated ovarian cancer cells.

Published
2018
Full Text
View/download PDF

10. Repurposing Pan-HDAC Inhibitors for ARID1A-Mutated Ovarian Cancer

Author

Takeshi Fukumoto, Pyoung Hwa Park, Shuai Wu, Nail Fatkhutdinov, Sergey Karakashev, Timothy Nacarelli, Andrew V. Kossenkov, David W. Speicher, Stephanie Jean, Lin Zhang, Tian-Li Wang, Ie-Ming Shih, Jose R. Conejo-Garcia, Benjamin G. Bitler, and Rugang Zhang

Subjects
Biology (General), QH301-705.5
Abstract

Summary: ARID1A, a subunit of the SWI/SNF complex, is among the most frequently mutated genes across cancer types. ARID1A is mutated in more than 50% of ovarian clear cell carcinomas (OCCCs), diseases that have no effective therapy. Here, we show that ARID1A mutation confers sensitivity to pan-HDAC inhibitors such as SAHA in ovarian cancers. This correlated with enhanced growth suppression induced by the inhibition of HDAC2 activity in ARID1A-mutated cells. HDAC2 interacts with EZH2 in an ARID1A status-dependent manner. HDAC2 functions as a co-repressor of EZH2 to suppress the expression of EZH2/ARID1A target tumor suppressor genes such as PIK3IP1 to inhibit proliferation and promote apoptosis. SAHA reduced the growth and ascites of the ARID1A-inactivated OCCCs in both orthotopic and genetic mouse models. This correlated with a significant improvement of survival of mice bearing ARID1A-mutated OCCCs. These findings provided preclinical rationales for repurposing FDA-approved pan-HDAC inhibitors for treating ARID1A-mutated cancers. : Fukumoto et al. show that ARID1A mutation confers sensitivity to pan-HDAC inhibitors such as SAHA in ovarian cancers. This correlated with enhanced growth suppression induced by the inhibition of HDAC2 activity in ARID1A-mutated cells. These findings provided preclinical rationales for repurposing FDA-approved pan-HDAC inhibitors for treating ARID1A-mutated cancers. Keywords: ovarian cancer, ARID1A, HDAC2, pan-HDAC inhibitor, SAHA, SWI/SNF, chromatin remodeling

Published
2018
Full Text
View/download PDF

11. CARM1-expressing ovarian cancer depends on the histone methyltransferase EZH2 activity

Author

Sergey Karakashev, Hengrui Zhu, Shuai Wu, Yuhki Yokoyama, Benjamin G. Bitler, Pyoung-Hwa Park, Jeong-Heon Lee, Andrew V. Kossenkov, Krutika Satish Gaonkar, Huihuang Yan, Ronny Drapkin, Jose R. Conejo-Garcia, David W. Speicher, Tamas Ordog, and Rugang Zhang

Subjects
Science
Abstract

CARM1 is an arginine methyltransferase often overexpressed in human cancer. Here, the authors show that EZH2 inhibition suppresses growth in CARM1-expressing epithelial ovarian cancer, and examine the mechanism of how CARM1 promotes EZH2-mediated tumor suppressor gene silencing.

Published
2018
Full Text
View/download PDF

12. Oxidation of Peroxiredoxin 6 in the Presence of GSH Increases its Phospholipase A2 Activity at Cytoplasmic pH

Author

Suiping Zhou, Chandra Dodia, Sheldon I. Feinstein, Sandra Harper, Henry J. Forman, David W. Speicher, and Aron B. Fisher

Subjects
substrate binding, sulfinic acid, Prdx6 structure, mass spectroscopic analysis, Therapeutics. Pharmacology, RM1-950
Abstract

The expression of the phospholipase A2 activity (aiPLA2) of peroxiredoxin 6 (Prdx6) in the cell cytoplasm is physiologically relevant for the repair of peroxidized cell membranes, but aiPLA2 assay in vitro indicates that, unlike assay at pH 4, activity at cytosolic pH is essentially absent with non-oxidized substrate. However, the addition of glutathione (GSH) to the assay medium significantly increased aiPLA2 activity at cytosolic pH, while oxidized GSH (GSSG) and several other thiols had no effect. By mass spectroscopy (ESI MS), the addition of GSH to Prdx6 paradoxically led to oxidation of its conserved Cys47 residue to a sulfinic acid. The effect of GSH on PLA2 activity was abolished by incubation under anaerobic conditions, confirming that auto-oxidation of the protein was the mechanism for the GSH effect. Analysis by circular dichroism (CD) and tryptophan fluorescence showed alterations of the protein structure in the presence of GSH. Independently of GSH, the oxidation of Prdx6 by exposure to H2O2 or the presence of oxidized phospholipid as substrate also significantly increased aiPLA2 activity at pH 7. We conclude that the oxidation of the peroxidatically active Cys47 of Prdx6 results in an increase of aiPLA2 activity at pH 7 without effect on the activity of the enzyme at pH 4.

Published
2018
Full Text
View/download PDF

13. Concurrent MEK2 Mutation and BRAF Amplification Confer Resistance to BRAF and MEK Inhibitors in Melanoma

Author

Jessie Villanueva, Jeffrey R. Infante, Clemens Krepler, Patricia Reyes-Uribe, Minu Samanta, Hsin-Yi Chen, Bin Li, Rolf K. Swoboda, Melissa Wilson, Adina Vultur, Mizuho Fukunaba-Kalabis, Bradley Wubbenhorst, Thomas Y. Chen, Qin Liu, Katrin Sproesser, Douglas J. DeMarini, Tona M. Gilmer, Anne-Marie Martin, Ronen Marmorstein, David C. Schultz, David W. Speicher, Giorgos C. Karakousis, Wei Xu, Ravi K. Amaravadi, Xiaowei Xu, Lynn M. Schuchter, Meenhard Herlyn, and Katherine L. Nathanson

Subjects
Biology (General), QH301-705.5
Abstract

Although BRAF and MEK inhibitors have proven clinical benefits in melanoma, most patients develop resistance. We report a de novo MEK2-Q60P mutation and BRAF gain in a melanoma from a patient who progressed on the MEK inhibitor trametinib and did not respond to the BRAF inhibitor dabrafenib. We also identified the same MEK2-Q60P mutation along with BRAF amplification in a xenograft tumor derived from a second melanoma patient resistant to the combination of dabrafenib and trametinib. Melanoma cells chronically exposed to trametinib acquired concurrent MEK2-Q60P mutation and BRAF-V600E amplification, which conferred resistance to MEK and BRAF inhibitors. The resistant cells had sustained MAPK activation and persistent phosphorylation of S6K. A triple combination of dabrafenib, trametinib, and the PI3K/mTOR inhibitor GSK2126458 led to sustained tumor growth inhibition. Hence, concurrent genetic events that sustain MAPK signaling can underlie resistance to both BRAF and MEK inhibitors, requiring novel therapeutic strategies to overcome it.

Published
2013
Full Text
View/download PDF

14. Targeting UGCG Overcomes Resistance to Lysosomal Autophagy Inhibition

Author

Vaibhav Jain, Sandra L. Harper, Amanda M. Versace, Dylan Fingerman, Gregory Schuyler Brown, Monika Bhardwaj, Mary Ann S. Crissey, Aaron R. Goldman, Gordon Ruthel, Qin Liu, Aleksandra Zivkovic, Holgar Stark, Meenhard Herlyn, Phyllis A. Gimotty, David W. Speicher, and Ravi K. Amaravadi

Subjects
Oncology
Abstract

Lysosomal autophagy inhibition (LAI) with hydroxychloroquine or DC661 can enhance cancer therapy, but tumor regrowth is common. To elucidate LAI resistance, proteomics and immunoblotting demonstrated that LAI induced lipid metabolism enzymes in multiple cancer cell lines. Lipidomics showed that LAI increased cholesterol, sphingolipids, and glycosphingolipids. These changes were associated with striking levels of GM1+ membrane microdomains (GMM) in plasma membranes and lysosomes. Inhibition of cholesterol/sphingolipid metabolism proteins enhanced LAI cytotoxicity. Targeting UDP-glucose ceramide glucosyltransferase (UGCG) synergistically augmented LAI cytotoxicity. Although UGCG inhibition decreased LAI-induced GMM and augmented cell death, UGCG overexpression led to LAI resistance. Melanoma patients with high UGCG expression had significantly shorter disease-specific survival. The FDA-approved UGCG inhibitor eliglustat combined with LAI significantly inhibited tumor growth and improved survival in syngeneic tumors and a therapy-resistant patient-derived xenograft. These findings nominate UGCG as a new cancer target, and clinical trials testing UGCG inhibition in combination with LAI are warranted. Significance: We discovered UGCG-dependent lipid remodeling drives resistance to LAI. Targeting UGCG with a drug approved for a lysosomal storage disorder enhanced LAI antitumor activity without toxicity. LAI and UGCG inhibition could be tested clinically in multiple cancers. This article is highlighted in the In This Issue feature, p. 247

Published
2022

15. Stromal changes in the aged lung induce an emergence from melanoma dormancy

Author

Mitchell E. Fane, Yash Chhabra, Gretchen M. Alicea, Devon A. Maranto, Stephen M. Douglass, Marie R. Webster, Vito W. Rebecca, Gloria E. Marino, Filipe Almeida, Brett L. Ecker, Daniel J. Zabransky, Laura Hüser, Thomas Beer, Hsin-Yao Tang, Andrew Kossenkov, Meenhard Herlyn, David W. Speicher, Wei Xu, Xiaowei Xu, Elizabeth M. Jaffee, Julio A. Aguirre-Ghiso, and Ashani T. Weeraratna

Subjects
Aging, Neoplasm, Residual, Multidisciplinary, c-Mer Tyrosine Kinase, Receptor Protein-Tyrosine Kinases, Fibroblasts, Axl Receptor Tyrosine Kinase, Article, Wnt-5a Protein, Proto-Oncogene Proteins, Tumor Microenvironment, Humans, Neoplasm Invasiveness, Neoplasm Metastasis, Stromal Cells, Lung, Melanoma, Aged, Skin
Abstract

Disseminated cancer cells from primary tumours can seed in distal tissues, but may take several years to form overt metastases, a phenomenon that is termed tumour dormancy. Despite its importance in metastasis and residual disease, few studies have been able to successfully characterize dormancy within melanoma. Here we show that the aged lung microenvironment facilitates a permissive niche for efficient outgrowth of dormant disseminated cancer cells—in contrast to the aged skin, in which age-related changes suppress melanoma growth but drive dissemination. These microenvironmental complexities can be explained by the phenotype switching model, which argues that melanoma cells switch between a proliferative cell state and a slower-cycling, invasive state(1–3). It was previously shown that dermal fibroblasts promote phenotype switching in melanoma during ageing(4–8). We now identify WNT5A as an activator of dormancy in melanoma disseminated cancer cells within the lung, which initially enables the efficient dissemination and seeding of melanoma cells in metastatic niches. Age-induced reprogramming of lung fibroblasts increases their secretion of the soluble WNT antagonist sFRP1, which inhibits WNT5A in melanoma cells and thereby enables efficient metastatic outgrowth. We also identify the tyrosine kinase receptors AXL and MER as promoting a dormancy-to-reactivation axis within melanoma cells. Overall, we find that age-induced changes in distal metastatic microenvironments promote the efficient reactivation of dormant melanoma cells in the lung.

Published
2022

16. Supplementary Data from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Supplementary Figures and Legends.

Published
2023

17. Supplementary Movie 1 from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Young BODIPY-C12 fibroblasts with GFP-Tagged melanoma cells showing lipid uptake

Published
2023

18. Supplementary Movie 2 from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Aged BODIPY-C12 fibroblasts with GFP-tagged melanoma cells

Published
2023

19. Supplementary Data Tables S1-S2 from Targeting UGCG Overcomes Resistance to Lysosomal Autophagy Inhibition

Author

Ravi K. Amaravadi, David W. Speicher, Phyllis A. Gimotty, Meenhard Herlyn, Holgar Stark, Aleksandra Zivkovic, Qin Liu, Gordon Ruthel, Aaron R. Goldman, Mary Ann S. Crissey, Monika Bhardwaj, Gregory Schuyler Brown, Dylan Fingerman, Amanda M. Versace, Sandra L. Harper, and Vaibhav Jain

Abstract

Supplemental Table S1: Patient Characteristics by UGCG expression defined Risk Groups in the Learning Dataset; Supplemental Table S2: Patient Characteristics by UGCG expression defined Risk Groups in the Validation Dataset (Cirenajwis et al., 2015)

Published
2023

20. Supplementary Data Figures S1-S5 from Targeting UGCG Overcomes Resistance to Lysosomal Autophagy Inhibition

Author

Ravi K. Amaravadi, David W. Speicher, Phyllis A. Gimotty, Meenhard Herlyn, Holgar Stark, Aleksandra Zivkovic, Qin Liu, Gordon Ruthel, Aaron R. Goldman, Mary Ann S. Crissey, Monika Bhardwaj, Gregory Schuyler Brown, Dylan Fingerman, Amanda M. Versace, Sandra L. Harper, and Vaibhav Jain

Abstract

Supplemental Figure S1: LAI remodels the lipidome of melanoma cells. Supplemental Figure S2: LAI induces UGCG, and UGCG inhibition augments DC661 cytotoxicity, and this effect is not seen by blocking cholesterol synthesis and key earlier steps of autophagy. Supplemental Figure S3: LAI increases the formation of GMM in cancer cells. Supplemental Figure S4: UGCG inhibition synergistically augments LAI induced cytotoxicity and abrogates GMM formation without elevating ceramide levels. Supplemental Figure S5: LAI induces UGCG-associated GMM formation in PDX cells, a combination of DC661 and eliglustat impairs tumor growth in a therapy-resistant PDX tumor model.

Published
2023

21. Data from Targeting UGCG Overcomes Resistance to Lysosomal Autophagy Inhibition

Author

Ravi K. Amaravadi, David W. Speicher, Phyllis A. Gimotty, Meenhard Herlyn, Holgar Stark, Aleksandra Zivkovic, Qin Liu, Gordon Ruthel, Aaron R. Goldman, Mary Ann S. Crissey, Monika Bhardwaj, Gregory Schuyler Brown, Dylan Fingerman, Amanda M. Versace, Sandra L. Harper, and Vaibhav Jain

Abstract

Lysosomal autophagy inhibition (LAI) with hydroxychloroquine or DC661 can enhance cancer therapy, but tumor regrowth is common. To elucidate LAI resistance, proteomics and immunoblotting demonstrated that LAI induced lipid metabolism enzymes in multiple cancer cell lines. Lipidomics showed that LAI increased cholesterol, sphingolipids, and glycosphingolipids. These changes were associated with striking levels of GM1+ membrane microdomains (GMM) in plasma membranes and lysosomes. Inhibition of cholesterol/sphingolipid metabolism proteins enhanced LAI cytotoxicity. Targeting UDP-glucose ceramide glucosyltransferase (UGCG) synergistically augmented LAI cytotoxicity. Although UGCG inhibition decreased LAI-induced GMM and augmented cell death, UGCG overexpression led to LAI resistance. Melanoma patients with high UGCG expression had significantly shorter disease-specific survival. The FDA-approved UGCG inhibitor eliglustat combined with LAI significantly inhibited tumor growth and improved survival in syngeneic tumors and a therapy-resistant patient-derived xenograft. These findings nominate UGCG as a new cancer target, and clinical trials testing UGCG inhibition in combination with LAI are warranted.Significance:We discovered UGCG-dependent lipid remodeling drives resistance to LAI. Targeting UGCG with a drug approved for a lysosomal storage disorder enhanced LAI antitumor activity without toxicity. LAI and UGCG inhibition could be tested clinically in multiple cancers.This article is highlighted in the In This Issue feature, p. 247

Published
2023

22. Supplementary Movie legends from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplemental Legends for Movies

Published
2023

23. Supplementary Movie 1 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 1

Published
2023

24. Supplementary Movie 2 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 2

Published
2023

25. Data from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Older patients with melanoma (>50 years old) have poorer prognoses and response rates to targeted therapy compared with young patients (Significance:These data show that melanoma cells take up lipids from aged fibroblasts, via FATP2, and use them to resist targeted therapy. The response to targeted therapy is altered in aged individuals because of the influences of the aged microenvironment, and these data suggest FATP2 as a target to overcome resistance.See related commentary by Montal and White, p. 1255..This article is highlighted in the In This Issue feature, p. 1241

Published
2023

26. Supplementary Movie 3 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 3

Published
2023

27. Supplementary Figures from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Figures

Published
2023

28. Chemical Methods from A Unified Approach to Targeting the Lysosome's Degradative and Growth Signaling Roles

Author

Ravi K. Amaravadi, Jeffrey D. Winkler, David W. Speicher, Donita C. Brady, Roberto Zoncu, Meenhard Herlyn, Xiaowei Xu, Shujing Liu, Qin Liu, Ronen Marmorstein, Joshua Rabinowitz, Yiling Lu, Gordon B. Mills, Maureen E. Murphy, Julie S. Barber-Rotenberg, Zhi Wei, Samuel M. Levi, Rani Ojha, Sengottuvelan Murugan, Shengfu Piao, Gretchen M. Alicea, Gao Zhang, Cynthia I. Chude, Eric Witze, Chun-Yan Lim, Michel Nofal, Amruta Ronghe, Quentin McAfee, Colin Fennelly, Noel McLaughlin, Michael C. Nicastri, and Vito W. Rebecca

Abstract

Supplemental Chemical Methods

Published
2023

29. Supplementary Movie 5 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 5

Published
2023

30. Supplementary Movie 8 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 8

Published
2023

31. Supplementary Movie 4 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 4

Published
2023

32. Source Code from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Source Code for analyses

Published
2023

33. Supplementary Movie 6 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 6

Published
2023

34. Supplementary Tables 1 and 2 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Tables of proteomics and CD4/8 staining

Published
2023

35. Supplementary Movie 2 (SM2) from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Aged BODIPY-C12 fibroblasts with GFP-tagged melanoma cells

Published
2023

36. Supplemental Figures S1-S7 and Supplemental Table S1 from A Unified Approach to Targeting the Lysosome's Degradative and Growth Signaling Roles

Author

Ravi K. Amaravadi, Jeffrey D. Winkler, David W. Speicher, Donita C. Brady, Roberto Zoncu, Meenhard Herlyn, Xiaowei Xu, Shujing Liu, Qin Liu, Ronen Marmorstein, Joshua Rabinowitz, Yiling Lu, Gordon B. Mills, Maureen E. Murphy, Julie S. Barber-Rotenberg, Zhi Wei, Samuel M. Levi, Rani Ojha, Sengottuvelan Murugan, Shengfu Piao, Gretchen M. Alicea, Gao Zhang, Cynthia I. Chude, Eric Witze, Chun-Yan Lim, Michel Nofal, Amruta Ronghe, Quentin McAfee, Colin Fennelly, Noel McLaughlin, Michael C. Nicastri, and Vito W. Rebecca

Abstract

Supplemental Figures and Table

Published
2023

37. Supplementary Figures S1-S4 from PPT1 Promotes Tumor Growth and Is the Molecular Target of Chloroquine Derivatives in Cancer

Author

Ravi K. Amaravadi, Jeffrey D. Winkler, David W. Speicher, Phyllis A. Gimotty, Ronen Marmorstein, Meenhard Herlyn, Xiaowei Xu, Lynn M. Schuchter, Jennifer J. Lee, Gretchen M. Alicea, Alessandra Martorella, Estela Noguera-Ortega, Shengfu Piao, Rani Ojha, Aaron R. Goldman, Gao Zhang, Noel P. McLaughlin, Quentin McAfee, Amruta Ronghe, Julie S. Barber-Rotenberg, Cynthia I. Chude, Colin Fennelly, Michael C. Nicastri, and Vito W. Rebecca

Abstract

Supplementary Figures S1-S4

Published
2023

38. Chemical Methods from PPT1 Promotes Tumor Growth and Is the Molecular Target of Chloroquine Derivatives in Cancer

Author

Ravi K. Amaravadi, Jeffrey D. Winkler, David W. Speicher, Phyllis A. Gimotty, Ronen Marmorstein, Meenhard Herlyn, Xiaowei Xu, Lynn M. Schuchter, Jennifer J. Lee, Gretchen M. Alicea, Alessandra Martorella, Estela Noguera-Ortega, Shengfu Piao, Rani Ojha, Aaron R. Goldman, Gao Zhang, Noel P. McLaughlin, Quentin McAfee, Amruta Ronghe, Julie S. Barber-Rotenberg, Cynthia I. Chude, Colin Fennelly, Michael C. Nicastri, and Vito W. Rebecca

Abstract

Chemical Methods

Published
2023

39. Supplementary Figure Legends from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Figure Legends

Published
2023

40. Supplementary Movie 1 (SM1) from Changes in Aged Fibroblast Lipid Metabolism Induce Age-Dependent Melanoma Cell Resistance to Targeted Therapy via the Fatty Acid Transporter FATP2

Author

Ashani T. Weeraratna, David W. Speicher, Zachary T. Schug, Ian A. Blair, Meenhard Herlyn, Dmitry I. Gabrilovich, Qin Liu, Xiaowei Xu, Keith T. Flaherty, Dennie T. Frederick, Hsin-Yao Tang, Andrew V. Kossenkov, M. Cecilia Caino, Brett L. Ecker, Curtis H. Kugel, Marie R. Webster, Reeti Behera, Stephen M. Douglass, Mitchell E. Fane, Aaron R. Goldman, Vito W. Rebecca, and Gretchen M. Alicea

Abstract

Young BODIPY-C12 fibroblasts with GFP-Tagged melanoma cells showing lipid uptake

Published
2023

41. Data from PPT1 Promotes Tumor Growth and Is the Molecular Target of Chloroquine Derivatives in Cancer

Author

Ravi K. Amaravadi, Jeffrey D. Winkler, David W. Speicher, Phyllis A. Gimotty, Ronen Marmorstein, Meenhard Herlyn, Xiaowei Xu, Lynn M. Schuchter, Jennifer J. Lee, Gretchen M. Alicea, Alessandra Martorella, Estela Noguera-Ortega, Shengfu Piao, Rani Ojha, Aaron R. Goldman, Gao Zhang, Noel P. McLaughlin, Quentin McAfee, Amruta Ronghe, Julie S. Barber-Rotenberg, Cynthia I. Chude, Colin Fennelly, Michael C. Nicastri, and Vito W. Rebecca

Abstract

Clinical trials repurposing lysosomotropic chloroquine (CQ) derivatives as autophagy inhibitors in cancer demonstrate encouraging results, but the underlying mechanism of action remains unknown. Here, we report a novel dimeric CQ (DC661) capable of deacidifying the lysosome and inhibiting autophagy significantly better than hydroxychloroquine (HCQ). Using an in situ photoaffinity pulldown strategy, we identified palmitoyl-protein thioesterase 1 (PPT1) as a molecular target shared across monomeric and dimeric CQ derivatives. HCQ and Lys05 also bound to and inhibited PPT1 activity, but only DC661 maintained activity in acidic media. Knockout of PPT1 in cancer cells using CRISPR/Cas9 editing abrogates autophagy modulation and cytotoxicity of CQ derivatives, and results in significant impairment of tumor growth similar to that observed with DC661. Elevated expression of PPT1 in tumors correlates with poor survival in patients in a variety of cancers. Thus, PPT1 represents a new target in cancer that can be inhibited with CQ derivatives.Significance:This study identifies PPT1 as the previously unknown lysosomal molecular target of monomeric and dimeric CQ derivatives. Genetic suppression of PPT1 impairs tumor growth, and PPT1 levels are elevated in cancer and associated with poor survival. These findings provide a strong rationale for targeting PPT1 in cancer.This article is highlighted in the In This Issue feature, p. 151

Published
2023

42. Supplementary Movie 9 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 9

Published
2023

43. Supplementary Movie 7 from Remodeling of the Collagen Matrix in Aging Skin Promotes Melanoma Metastasis and Affects Immune Cell Motility

Author

Ashani T. Weeraratna, Edna Cukierman, Vivek Shenoy, Arjun Raj, Lauren E. Haydu, Michael T. Tetzlaff, Jennifer A. Wargo, David W. Speicher, Meenhard Herlyn, Patricia Brafford, Mitchell Fane, Xiaowei Xu, Qin Liu, Xiangfan Yin, Hsin-Yao Tang, Andrew Kossenkov, Frederick Keeney, Ian A. Mellis, Neelima Shah, Janusz Franco-Barraza, Hossein Ahmadzadeh, Ehsan Ban, James Hayden, Rajasekharan Somasundaram, Filipe V. Almeida, Marie R. Webster, Curtis H. Kugel, Stephen M. Douglass, Brett L. Ecker, and Amanpreet Kaur

Abstract

Supplementary Movie 7

Published
2023

44. Table S3 from Neural Crest-Like Stem Cell Transcriptome Analysis Identifies LPAR1 in Melanoma Progression and Therapy Resistance

Author

Meenhard Herlyn, Ze'ev A. Ronai, Keith T. Flaherty, David W. Speicher, Joseph M. Salvino, Genevieve M. Boland, Dennie T. Frederick, Lawrence N. Kwong, Ravi K. Amaravadi, Tara C. Mitchell, Lynn M. Schuchter, Xiaowei Xu, Giorgos C. Karakousis, Wei Xu, Gordon B. Mills, Yiling Lu, Lawrence W. Wu, Denitsa M. Hristova, Adina Vultur, Rajasekharan Somasundaram, Marilda Beqiri, Joshua X. Wang, Patricia Brafford, Katrin Sproesser, Alice S. Morias, Maya Delvadia, Ye Huang, Cynthia Tong, Bela Delvadia, Chih-Cheng Yang, Pedro Aza-Blanc, Clemens Krepler, Mizuho Fukunaga-Kalabis, Gretchen M. Alicea, Zhi Wei, Chaoran Cheng, Jie Zhang, Delaine Zayasbazan, Anastasia Samarkina, Gao Zhang, Ling Li, Min Xiao, Alexis Gutierrez, Qin Liu, Thomas Connelly, Andrew V. Kossenkov, Vito W. Rebecca, and Jianglan Liu

Abstract

Supplementary Table

Published
2023

45. Data from Neural Crest-Like Stem Cell Transcriptome Analysis Identifies LPAR1 in Melanoma Progression and Therapy Resistance

Author

Meenhard Herlyn, Ze'ev A. Ronai, Keith T. Flaherty, David W. Speicher, Joseph M. Salvino, Genevieve M. Boland, Dennie T. Frederick, Lawrence N. Kwong, Ravi K. Amaravadi, Tara C. Mitchell, Lynn M. Schuchter, Xiaowei Xu, Giorgos C. Karakousis, Wei Xu, Gordon B. Mills, Yiling Lu, Lawrence W. Wu, Denitsa M. Hristova, Adina Vultur, Rajasekharan Somasundaram, Marilda Beqiri, Joshua X. Wang, Patricia Brafford, Katrin Sproesser, Alice S. Morias, Maya Delvadia, Ye Huang, Cynthia Tong, Bela Delvadia, Chih-Cheng Yang, Pedro Aza-Blanc, Clemens Krepler, Mizuho Fukunaga-Kalabis, Gretchen M. Alicea, Zhi Wei, Chaoran Cheng, Jie Zhang, Delaine Zayasbazan, Anastasia Samarkina, Gao Zhang, Ling Li, Min Xiao, Alexis Gutierrez, Qin Liu, Thomas Connelly, Andrew V. Kossenkov, Vito W. Rebecca, and Jianglan Liu

Abstract

Metastatic melanoma is challenging to clinically address. Although standard-of-care targeted therapy has high response rates in patients with BRAF-mutant melanoma, therapy relapse occurs in most cases. Intrinsically resistant melanoma cells drive therapy resistance and display molecular and biologic properties akin to neural crest-like stem cells (NCLSC) including high invasiveness, plasticity, and self-renewal capacity. The shared transcriptional programs and vulnerabilities between NCLSCs and cancer cells remains poorly understood. Here, we identify a developmental LPAR1-axis critical for NCLSC viability and melanoma cell survival. LPAR1 activity increased during progression and following acquisition of therapeutic resistance. Notably, genetic inhibition of LPAR1 potentiated BRAFi ± MEKi efficacy and ablated melanoma migration and invasion. Our data define LPAR1 as a new therapeutic target in melanoma and highlights the promise of dissecting stem cell–like pathways hijacked by tumor cells.Significance:This study identifies an LPAR1-axis critical for melanoma invasion and intrinsic/acquired therapy resistance.

Published
2023

46. Supplementary Figures from Neural Crest-Like Stem Cell Transcriptome Analysis Identifies LPAR1 in Melanoma Progression and Therapy Resistance

Author

Meenhard Herlyn, Ze'ev A. Ronai, Keith T. Flaherty, David W. Speicher, Joseph M. Salvino, Genevieve M. Boland, Dennie T. Frederick, Lawrence N. Kwong, Ravi K. Amaravadi, Tara C. Mitchell, Lynn M. Schuchter, Xiaowei Xu, Giorgos C. Karakousis, Wei Xu, Gordon B. Mills, Yiling Lu, Lawrence W. Wu, Denitsa M. Hristova, Adina Vultur, Rajasekharan Somasundaram, Marilda Beqiri, Joshua X. Wang, Patricia Brafford, Katrin Sproesser, Alice S. Morias, Maya Delvadia, Ye Huang, Cynthia Tong, Bela Delvadia, Chih-Cheng Yang, Pedro Aza-Blanc, Clemens Krepler, Mizuho Fukunaga-Kalabis, Gretchen M. Alicea, Zhi Wei, Chaoran Cheng, Jie Zhang, Delaine Zayasbazan, Anastasia Samarkina, Gao Zhang, Ling Li, Min Xiao, Alexis Gutierrez, Qin Liu, Thomas Connelly, Andrew V. Kossenkov, Vito W. Rebecca, and Jianglan Liu

Abstract

Supplementary Figures

Published
2023

48. Figure S6 from MFF Regulation of Mitochondrial Cell Death Is a Therapeutic Target in Cancer

Author

Dario C. Altieri, Emmanuel Skordalakes, Alessandra Martorella, Dylan Fingerman, Min Xiao, Meenhard Herlyn, Vito W. Rebecca, Valentina Vaira, Massimo Giroda, Davide Tosi, Manuela Caroli, Gabriella Gaudioso, Stefano Ferrero, Alessandra Maria Storaci, Prashanth K. Shastrula, David W. Speicher, Lucia R. Languino, Dmitry I. Gabrilovich, Ekta Agarwal, Hsin-Yao Tang, Yu Geon Lee, Andrew V. Kossenkov, Young Chan Chae, and Jae Ho Seo

Abstract

Anti-glioma activity of MFF peptidomimetic

Published
2023

49. Data from A Novel Inhibitor of HSP70 Induces Mitochondrial Toxicity and Immune Cell Recruitment in Tumors

Author

Maureen E. Murphy, Donna L. George, Joseph M. Salvino, Dmitry I. Gabrilovich, David W. Speicher, Qin Liu, Andrew V. Kossenkov, Joel Cassel, Hsin-Yao Tang, Rebecca Moeller, Emilio Sanseviero, Keerthana Gnanapradeepan, Madeline Good, Tetyana Martynyuk, Alexandra Indeglia, Joshua L.D. Parris, Adi Narayana Reddy Poli, Subhasree Basu, Julia I-Ju Leu, Jessica C. Leung, and Thibaut Barnoud

Abstract

The protein chaperone HSP70 is overexpressed in many cancers including colorectal cancer, where overexpression is associated with poor survival. We report here the creation of a uniquely acting HSP70 inhibitor (HSP70i) that targets multiple compartments in the cancer cell, including mitochondria. This inhibitor was mitochondria toxic and cytotoxic to colorectal cancer cells, but not to normal colon epithelial cells. Inhibition of HSP70 was efficacious as a single agent in primary and metastatic models of colorectal cancer and enabled identification of novel mitochondrial client proteins for HSP70. In a syngeneic colorectal cancer model, the inhibitor increased immune cell recruitment into tumors. Cells treated with the inhibitor secreted danger-associated molecular patterns (DAMP), including ATP and HMGB1, and functioned effectively as a tumor vaccine. Interestingly, the unique properties of this HSP70i in the disruption of mitochondrial function and the inhibition of proteostasis both contributed to DAMP release. This HSP70i constitutes a promising therapeutic opportunity in colorectal cancer and may exhibit antitumor activity against other tumor types.Significance:These findings describe a novel HSP70i that disrupts mitochondrial proteostasis, demonstrating single-agent efficacy that induces immunogenic cell death in treated tumors.

Published
2023

Catalog

Books, media, physical & digital resources
See catalog results