27 results on '"Kiran Kurmi"'
Search Results
2. Carnitine Palmitoyltransferase 1A Has a Lysine Succinyltransferase Activity
- Author
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Kiran Kurmi, Sadae Hitosugi, Elizabeth K. Wiese, Felix Boakye-Agyeman, Wilson I. Gonsalves, Zhenkun Lou, Larry M. Karnitz, Matthew P. Goetz, and Taro Hitosugi
- Subjects
post-translational modification ,signal transduction ,metabolism ,lysine succinyltransferase ,lysine succinylation ,carnitine palmitoyltransferase 1A ,Biology (General) ,QH301-705.5 - Abstract
Summary: Lysine succinylation was recently identified as a post-translational modification in cells. However, the molecular mechanism underlying lysine succinylation remains unclear. Here, we show that carnitine palmitoyltransferase 1A (CPT1A) has lysine succinyltransferase (LSTase) activity in vivo and in vitro. Using a stable isotope labeling by amino acid in cell culture (SILAC)-based proteomics approach, we found that 101 proteins were more succinylated in cells expressing wild-type (WT) CPT1A compared with vector control cells. One of the most heavily succinylated proteins in this analysis was enolase 1. We found that CPT1A WT succinylated enolase 1 and reduced enolase enzymatic activity in cells and in vitro. Importantly, mutation of CPT1A Gly710 (G710E) selectively inactivated carnitine palmitoyltransferase (CPTase) activity but not the LSTase activity that decreased enolase activity in cells and promoted cell proliferation under glutamine depletion. These findings suggest that CPT1A acts as an LSTase that can regulate enzymatic activity of a substrate protein and metabolism independent of its classical CPTase activity.
- Published
- 2018
- Full Text
- View/download PDF
3. Supplementary Figure and Table Legends from Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Arlene H. Sharpe, Marcia C. Haigis, Justin D. Trombley, Vikram R. Juneja, Kiran Kurmi, Cong-Hui Yao, Alison E. Ringel, Martin W. LaFleur, Isaac S. Harris, Emily F. Gaudiano, Thao H. Nguyen, Tara Muijlwijk, Jacob E. Gillis, and Jefte M. Drijvers
- Abstract
Supplementary Figure and Table Legends
- Published
- 2023
4. Supplementary Figures from Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Arlene H. Sharpe, Marcia C. Haigis, Justin D. Trombley, Vikram R. Juneja, Kiran Kurmi, Cong-Hui Yao, Alison E. Ringel, Martin W. LaFleur, Isaac S. Harris, Emily F. Gaudiano, Thao H. Nguyen, Tara Muijlwijk, Jacob E. Gillis, and Jefte M. Drijvers
- Abstract
Supplementary Figures
- Published
- 2023
5. Supplementary Table S1 from Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Arlene H. Sharpe, Marcia C. Haigis, Justin D. Trombley, Vikram R. Juneja, Kiran Kurmi, Cong-Hui Yao, Alison E. Ringel, Martin W. LaFleur, Isaac S. Harris, Emily F. Gaudiano, Thao H. Nguyen, Tara Muijlwijk, Jacob E. Gillis, and Jefte M. Drijvers
- Abstract
Supplementary Table S1
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- 2023
6. Supplementary Data File 1 from Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Arlene H. Sharpe, Marcia C. Haigis, Justin D. Trombley, Vikram R. Juneja, Kiran Kurmi, Cong-Hui Yao, Alison E. Ringel, Martin W. LaFleur, Isaac S. Harris, Emily F. Gaudiano, Thao H. Nguyen, Tara Muijlwijk, Jacob E. Gillis, and Jefte M. Drijvers
- Abstract
All dose-response curves. Dose-response curves for all library compounds
- Published
- 2023
7. Data from Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Arlene H. Sharpe, Marcia C. Haigis, Justin D. Trombley, Vikram R. Juneja, Kiran Kurmi, Cong-Hui Yao, Alison E. Ringel, Martin W. LaFleur, Isaac S. Harris, Emily F. Gaudiano, Thao H. Nguyen, Tara Muijlwijk, Jacob E. Gillis, and Jefte M. Drijvers
- Abstract
Metabolic constraints in the tumor microenvironment constitute a barrier to effective antitumor immunity and similarities in the metabolic properties of T cells and cancer cells impede the specific therapeutic targeting of metabolism in either population. To identify distinct metabolic vulnerabilities of CD8+ T cells and cancer cells, we developed a high-throughput in vitro pharmacologic screening platform and used it to measure the cell type–specific sensitivities of activated CD8+ T cells and B16 melanoma cells to a wide array of metabolic perturbations during antigen-specific killing of cancer cells by CD8+ T cells. We illustrated the applicability of this screening platform by showing that CD8+ T cells were more sensitive to ferroptosis induction by inhibitors of glutathione peroxidase 4 (GPX4) than B16 and MC38 cancer cells. Overexpression of ferroptosis suppressor protein 1 (FSP1) or cytosolic GPX4 yielded ferroptosis-resistant CD8+ T cells without compromising their function, while genetic deletion of the ferroptosis sensitivity–promoting enzyme acyl-CoA synthetase long-chain family member 4 (ACSL4) protected CD8+ T cells from ferroptosis but impaired antitumor CD8+ T-cell responses. Our screen also revealed high T cell–specific vulnerabilities for compounds targeting NAD+ metabolism or autophagy and endoplasmic reticulum (ER) stress pathways. We focused the current screening effort on metabolic agents. However, this in vitro screening platform may also be valuable for rapid testing of other types of compounds to identify regulators of antitumor CD8+ T-cell function and potential therapeutic targets.
- Published
- 2023
8. Supplementary Table S2 from BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents
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Larry M. Karnitz, S. John Weroha, Scott H. Kaufmann, Taro Hitosugi, Ann L. Oberg, Hu Li, Sean C. Dowdy, Andrea E. Wahner Hendrickson, Daniel O'Brien, Catherine J. Huntoon, Cristina Correia, Ethan P. Heinzen, Emma R. Purfeerst, Xiaonan Hou, Thomas L. Ekstrom, Kiran Kurmi, and Arun Kanakkanthara
- Abstract
Excel data file of NNMT and CDK12 protein levels in HGSOC tumors from patients.
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- 2023
9. Supplementary Figures S1-6. from BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents
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Larry M. Karnitz, S. John Weroha, Scott H. Kaufmann, Taro Hitosugi, Ann L. Oberg, Hu Li, Sean C. Dowdy, Andrea E. Wahner Hendrickson, Daniel O'Brien, Catherine J. Huntoon, Cristina Correia, Ethan P. Heinzen, Emma R. Purfeerst, Xiaonan Hou, Thomas L. Ekstrom, Kiran Kurmi, and Arun Kanakkanthara
- Abstract
Supplementary Figures S1-6. Supplementary Fig. S1 shows that CDK12 depletion alters ATP and ADP levels without affecting glycolysis in cell lines. Supplementary Fig. S2 shows that BRCA1 depletion does not affect glycolysis, that CDK12 and BRCA1 depletion reduce ATP levels in ex vivo HGSOC PDX cultures, that transient BRCA1 expression increase OCR in BRCA1-deficient cells, that CDK12 and BRCA1 depletion do not affect mitochondrial DNA levels, and levels of BRCA1 and CDK12 mRNA expression in cells transfected with CDK12 siRNAs and BRCA1 expression plasmid. Supplementary Fig. S3 shows BRCA2 and RAD51 depletion do not affect OCR but do sensitize to olaparib. Supplementary Fig. S4 shows a map of known BRCA1 transcripts, the exons that are targeted by the BRCA1 siRNAs used in this study, primers used to assess expression of alternative BRCA1 transcripts, NNMT mRNA expression in cells transfected with BRCA1 siRNAs, and binding of BRCA1 to the NNMT promoter using ChIP. Supplementary S5 shows that BRCA2 and RAD51 depletion do not sensitize to VLX600 but do sensitize to olaparib. Supplementary Fig. S6 shows that CDK12 and BRCA1 depletion does not further suppress ATP levels in NNMT-overexpressing cells.
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- 2023
10. Supplementary Methods and oligonucleotide sequences from BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents
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Larry M. Karnitz, S. John Weroha, Scott H. Kaufmann, Taro Hitosugi, Ann L. Oberg, Hu Li, Sean C. Dowdy, Andrea E. Wahner Hendrickson, Daniel O'Brien, Catherine J. Huntoon, Cristina Correia, Ethan P. Heinzen, Emma R. Purfeerst, Xiaonan Hou, Thomas L. Ekstrom, Kiran Kurmi, and Arun Kanakkanthara
- Abstract
Supplementary Methods file that includes oligonucleotide sequences used in the study.
- Published
- 2023
11. Data from BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents
- Author
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Larry M. Karnitz, S. John Weroha, Scott H. Kaufmann, Taro Hitosugi, Ann L. Oberg, Hu Li, Sean C. Dowdy, Andrea E. Wahner Hendrickson, Daniel O'Brien, Catherine J. Huntoon, Cristina Correia, Ethan P. Heinzen, Emma R. Purfeerst, Xiaonan Hou, Thomas L. Ekstrom, Kiran Kurmi, and Arun Kanakkanthara
- Abstract
BRCA1 plays a key role in homologous recombination (HR) DNA repair. Accordingly, changes that downregulate BRCA1, including BRCA1 mutations and reduced BRCA1 transcription, due to promoter hypermethylation or loss of the BRCA1 transcriptional regulator CDK12, disrupt HR in multiple cancers. In addition, BRCA1 has also been implicated in the regulation of metabolism. Here, we show that reducing BRCA1 expression, either by CDK12 or BRCA1 depletion, led to metabolic reprogramming of ovarian cancer cells, causing decreased mitochondrial respiration and reduced ATP levels. BRCA1 depletion drove this reprogramming by upregulating nicotinamide N-methyltransferase (NNMT). Notably, the metabolic alterations caused by BRCA1 depletion and NNMT upregulation sensitized ovarian cancer cells to agents that inhibit mitochondrial metabolism (VLX600 and tigecycline) and to agents that inhibit glucose import (WZB117). These observations suggest that inhibition of energy metabolism may be a potential strategy to selectively target BRCA1-deficient high-grade serous ovarian cancer, which is characterized by frequent BRCA1 loss and NNMT overexpression.Significance:Loss of BRCA1 reprograms metabolism, creating a therapeutically targetable vulnerability in ovarian cancer.
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- 2023
12. Intercellular nanotubes mediate mitochondrial trafficking between cancer and immune cells
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Ruparoshni Jayabalan, Jayanta Mondal, Sachin K. Khiste, Chinmayee Dash, Hae Lin Jang, Tanmoy Saha, Kiran Kurmi, Pradip K. Majumder, Shiladitya Sengupta, Aditya Bardia, and Arpita Kulkarni
- Subjects
T-Lymphocytes ,Farnesyltransferase ,medicine.medical_treatment ,Biomedical Engineering ,Bioengineering ,Mitochondrion ,Article ,Immune system ,Neoplasms ,medicine ,General Materials Science ,Electrical and Electronic Engineering ,biology ,Chemistry ,Cancer ,Metabolism ,Immunotherapy ,biochemical phenomena, metabolism, and nutrition ,Condensed Matter Physics ,medicine.disease ,Atomic and Molecular Physics, and Optics ,Mitochondria ,Cell biology ,Cancer cell ,biology.protein ,bacteria ,Intracellular - Abstract
Cancer progresses by evading the immune system. Elucidating diverse immune evasion strategies is a critical step in the search for next-generation immunotherapies for cancer. Here we report that cancer cells can hijack the mitochondria from immune cells via physical nanotubes. Mitochondria are essential for metabolism and activation of immune cells. By using field-emission scanning electron microscopy, fluorophore-tagged mitochondrial transfer tracing and metabolic quantification, we demonstrate that the nanotube-mediated transfer of mitochondria from immune cells to cancer cells metabolically empowers the cancer cells and depletes the immune cells. Inhibiting the nanotube assembly machinery significantly reduced mitochondrial transfer and prevented the depletion of immune cells. Combining a farnesyltransferase and geranylgeranyltransferase 1 inhibitor, namely, L-778123, which partially inhibited nanotube formation and mitochondrial transfer, with a programmed cell death protein 1 immune checkpoint inhibitor improved the antitumour outcomes in an aggressive immunocompetent breast cancer model. Nanotube-mediated mitochondrial hijacking can emerge as a novel target for developing next-generation immunotherapy agents for cancer. Cancer cells adopt a series of strategies to evade the immune response mounted by the organism against them. Here we find that tumour cells can hijack mitochondria from immune cells by forming physical nanotubes, and suggest that inhibiting this process might represent a potential immunotherapy approach.
- Published
- 2021
13. Enzymatic activation of pyruvate kinase increases cytosolic oxaloacetate to inhibit the Warburg effect
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Wilson I. Gonsalves, Larry M. Karnitz, Sadae Hitosugi, Kiran Kurmi, Taro Hitosugi, Annapoorna Sreedhar, Elizabeth K. Wiese, Yuan-Ping Pang, Lindsey G. Andres-Beck, and Sharon T. Loa
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Oxaloacetic Acid ,Endocrinology, Diabetes and Metabolism ,Lactate dehydrogenase A ,Pyruvate Kinase ,PKM2 ,Article ,Mice ,Cytosol ,Cell Line, Tumor ,Physiology (medical) ,Internal Medicine ,Animals ,Humans ,Citrate synthase ,Glycolysis ,Enzyme Inhibitors ,education ,education.field_of_study ,Glutaminolysis ,biology ,Chemistry ,Activator (genetics) ,Cell Biology ,Warburg effect ,Enzyme Activation ,Glucose ,Gene Expression Regulation ,Biochemistry ,biology.protein ,Rabbits ,Lactate Dehydrogenase 5 ,Pyruvate kinase - Abstract
Pharmacological activation of the glycolytic enzyme PKM2 or expression of the constitutively active PKM1 isoform in cancer cells results in decreased lactate production, a phenomenon known as the PKM2 paradox in the Warburg effect. Here we show that oxaloacetate (OAA) is a competitive inhibitor of human lactate dehydrogenase A (LDHA) and that elevated PKM2 activity increases de novo synthesis of OAA through glutaminolysis, thereby inhibiting LDHA in cancer cells. We also show that replacement of human LDHA with rabbit LDHA, which is relatively resistant to OAA inhibition, eliminated the paradoxical correlation between the elevated PKM2 activity and the decreased lactate concentration in cancer cells treated with a PKM2 activator. Furthermore, rabbit LDHA-expressing tumours, compared to human LDHA-expressing tumours in mice, displayed resistance to the PKM2 activator. These findings describe a mechanistic explanation for the PKM2 paradox by showing that OAA accumulates and inhibits LDHA following PKM2 activation. Wiese et al. find that oxaloacetate generated through increased activation of PKM2 can inhibit lactate dehydrogenase A, shedding light on the long observed PKM2 paradox during Warburg metabolism in cancer cells.
- Published
- 2021
14. Oncometabolite d-2HG alters T cell metabolism to impair CD8
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Giulia Notarangelo, Jessica B. Spinelli, Elizabeth M. Perez, Gregory J. Baker, Kiran Kurmi, Ilaria Elia, Sylwia A. Stopka, Gerard Baquer, Jia-Ren Lin, Alexandra J. Golby, Shakchhi Joshi, Heide F. Baron, Jefte M. Drijvers, Peter Georgiev, Alison E. Ringel, Elma Zaganjor, Samuel K. McBrayer, Peter K. Sorger, Arlene H. Sharpe, Kai W. Wucherpfennig, Sandro Santagata, Nathalie Y. R. Agar, Mario L. Suvà, and Marcia C. Haigis
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IDH ,Carcinogenesis ,T-Lymphocytes ,CD8-Positive T-Lymphocytes ,PHENOTYPE ,Article ,Glutarates ,Interferon-gamma ,Mice ,DEHYDROGENASE ,MITOCHONDRIA ,Neoplasms ,Animals ,Humans ,Science & Technology ,IFN-GAMMA ,Multidisciplinary ,2-HYDROXYGLUTARATE ,L-Lactate Dehydrogenase ,MUTATIONS ,MAGNETIC-RESONANCE-SPECTROSCOPY ,Isocitrate Dehydrogenase ,Multidisciplinary Sciences ,ALPHA ,DIFFERENTIATION ,Gain of Function Mutation ,Mutation ,Science & Technology - Other Topics - Abstract
Gain-of-function mutations in isocitrate dehydrogenase (IDH) in human cancers result in the production of d -2-hydroxyglutarate ( d -2HG), an oncometabolite that promotes tumorigenesis through epigenetic alterations. The cancer cell–intrinsic effects of d -2HG are well understood, but its tumor cell–nonautonomous roles remain poorly explored. We compared the oncometabolite d -2HG with its enantiomer, l -2HG, and found that tumor-derived d -2HG was taken up by CD8 + T cells and altered their metabolism and antitumor functions in an acute and reversible fashion. We identified the glycolytic enzyme lactate dehydrogenase (LDH) as a molecular target of d -2HG. d -2HG and inhibition of LDH drive a metabolic program and immune CD8 + T cell signature marked by decreased cytotoxicity and impaired interferon-γ signaling that was recapitulated in clinical samples from human patients with IDH1 mutant gliomas.
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- 2022
15. Abstract 1156: GUK1 is a novel metabolic liability in oncogene-driven lung cancer
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Jaime Laurel Schneider, Kiran Kurmi, Ishita Dhiman, Roberta Colapietro, Shakchhi Joshi, Christian Johnson, Satoshi Yoda, Joao Paulo, Daniela Ruiz, Sylwia Stopka, Gerard Baquer, Jessica Lin, Kevin Haigis, Nathalie Agar, Steven Gygi, Aaron Hata, and Marcia Haigis
- Subjects
Cancer Research ,Oncology - Abstract
There is a longstanding desire to take therapeutic advantage of dysregulated metabolic states in cancer. While it has been appreciated that lung tumors rewire their cellular metabolic networks to support unrestrained proliferation, metabolic vulnerabilities have largely not been explored in the context of specific onco-genotypes. This represents a major gap in our understanding of how different oncogenic drivers in non-small cell lung cancer (NSCLC) confer reliance on discrete metabolic networks to sustain tumor growth. The goals of this project are (1) to investigate metabolic dependencies in distinct molecular subtypes of lung cancer and (2) to elucidate how metabolic reprogramming drives resistance to targeted therapy. Using patient-derived cell culture models and tumor specimens collected from patients with ALK-positive (ALK+) NSCLC, we identified that lung tumors with ALK rearrangements harbor a unique metabolic signature marked by reliance on anabolic nucleotide pathways. A phosphoproteomic screen in ALK+ patient-derived cells identified a novel metabolic target of ALK signaling, GUK1, the only known enzyme responsible for GDP synthesis. We show that ALK binds to and phosphorylates GUK1 and that ALK-mediated GUK1 phosphorylation augments GDP/GTP nucleotide biosynthesis. Steady-state and tracing metabolomic studies demonstrate that ALK inhibition and GUK1 phosphomutant are epistatic in guanine nucleotide production. Molecular dynamic modeling suggests that phosphorylation of GUK1 alters the dynamics of active site closure to enhance substrate processivity and protects GUK1 from a non-catalytic confirmation. Introduction of phosphomutant GUK1 into ALK+ patient-derived cell lines results in decreased tumor proliferation in vitro and in vivo in xenograft models. Spatially resolved mass spectrometry imaging of tumor specimens from ALK+ patients demonstrates significant enrichment of guanine nucleotides in ALK+ and phospho-GUK1+ tumor cells. We identified that other oncogenic fusion proteins regulate GUK1 phosphorylation, highlighting the need to further characterize GUK1 as a metabolic liability in NSCLC. Furthermore, a subset of patient-derived cell lines with resistance to ALK tyrosine kinase inhibitors (TKIs) exhibits increased expression and phosphorylation of GUK1, indicating that regulation of this metabolic enzyme may play a role in mediating acquired resistance. We anticipate these studies will pave the way for the development of new therapeutic approaches by exploiting metabolic vulnerabilities in oncogene-driven lung cancers. Citation Format: Jaime Laurel Schneider, Kiran Kurmi, Ishita Dhiman, Roberta Colapietro, Shakchhi Joshi, Christian Johnson, Satoshi Yoda, Joao Paulo, Daniela Ruiz, Sylwia Stopka, Gerard Baquer, Jessica Lin, Kevin Haigis, Nathalie Agar, Steven Gygi, Aaron Hata, Marcia Haigis. GUK1 is a novel metabolic liability in oncogene-driven lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1156.
- Published
- 2023
16. Abstract 3680: Multiomics analysis of triple negative breast cancer identifies potential metabolic vulnerability for overcoming the resistance of neoadjuvant therapy
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Zuen Ren, Kiran Kurmi, Robert Morris, Shakchhi Joshi, Eric Zaniewski, Johannes Kreuzer, Gabrielle Elena Gioia, Wilhelm Haas, Marcia C. Haigis, and Leif W. Ellisen
- Subjects
Cancer Research ,Oncology - Abstract
Triple-negative breast cancer (TNBC) is a heterogeneous disease that remains currently medically incurable. We comprehensively analyzed clinical, proteomic, and metabolomic data of a cohort of 12 highly selected primary TNBCs. Among them, 7 patients after neoadjuvant chemotherapy had residual disease and micro- or macro-metastases that were non-pathologically complete response (non-pCR); 5 patients had pathologically complete response (pCR). Interestingly, both proteomic and metabolic profiling results revealed dramatically different molecular signatures in non-pCR relative to pCR. Non-pCR were enriched in metabolites involving aspartic acid, fumarate, nicotinamide, and adenosine that are critical intermediates for nitrogen recycling and tricarboxylic acid (TCA) cycle, which are essential fuels for tumor biomass. Moreover, non-pCR were enriched in urea cycle, oxidative phosphorylation, glycolysis, sirtuin signaling, nucleotide De Novo biosynthesis, which are resistance-associated. Putative therapeutic targets were identified in non-pCR patients. Collectively, our multiomics analysis demonstrated the heterogeneity of TNBCs at multi-levels and enabled the development of personalized therapies targeting unique tumor metabolic profiles. Citation Format: Zuen Ren, Kiran Kurmi, Robert Morris, Shakchhi Joshi, Eric Zaniewski, Johannes Kreuzer, Gabrielle Elena Gioia, Wilhelm Haas, Marcia C. Haigis, Leif W. Ellisen. Multiomics analysis of triple negative breast cancer identifies potential metabolic vulnerability for overcoming the resistance of neoadjuvant therapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3680.
- Published
- 2023
17. Uncoupled glycerol-3-phosphate shuttle in kidney cancer reveals that cytosolic GPD is essential to support lipid synthesis
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Cong-Hui Yao, Joon Seok Park, Kiran Kurmi, Song-Hua Hu, Giulia Notarangelo, Joseph Crowley, Heidi Jacobson, Sheng Hui, Arlene H. Sharpe, and Marcia C. Haigis
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Cell Biology ,Molecular Biology - Published
- 2023
18. Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
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Vikram R. Juneja, Arlene H. Sharpe, Isaac S. Harris, Jacob E. Gillis, Emily F. Gaudiano, Marcia C. Haigis, Thao H. Nguyen, Alison E. Ringel, Tara Muijlwijk, Jefte M. Drijvers, Martin W. LaFleur, Cong-Hui Yao, Kiran Kurmi, and Justin D. Trombley
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0301 basic medicine ,Cancer Research ,education.field_of_study ,Tumor microenvironment ,Chemistry ,Endoplasmic reticulum ,Immunology ,Population ,Cell ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer cell ,medicine ,Cancer research ,Cytotoxic T cell ,NAD+ kinase ,education ,CD8 - Abstract
Metabolic constraints in the tumor microenvironment constitute a barrier to effective antitumor immunity and similarities in the metabolic properties of T cells and cancer cells impede the specific therapeutic targeting of metabolism in either population. To identify distinct metabolic vulnerabilities of CD8+ T cells and cancer cells, we developed a high-throughput in vitro pharmacologic screening platform and used it to measure the cell type–specific sensitivities of activated CD8+ T cells and B16 melanoma cells to a wide array of metabolic perturbations during antigen-specific killing of cancer cells by CD8+ T cells. We illustrated the applicability of this screening platform by showing that CD8+ T cells were more sensitive to ferroptosis induction by inhibitors of glutathione peroxidase 4 (GPX4) than B16 and MC38 cancer cells. Overexpression of ferroptosis suppressor protein 1 (FSP1) or cytosolic GPX4 yielded ferroptosis-resistant CD8+ T cells without compromising their function, while genetic deletion of the ferroptosis sensitivity–promoting enzyme acyl-CoA synthetase long-chain family member 4 (ACSL4) protected CD8+ T cells from ferroptosis but impaired antitumor CD8+ T-cell responses. Our screen also revealed high T cell–specific vulnerabilities for compounds targeting NAD+ metabolism or autophagy and endoplasmic reticulum (ER) stress pathways. We focused the current screening effort on metabolic agents. However, this in vitro screening platform may also be valuable for rapid testing of other types of compounds to identify regulators of antitumor CD8+ T-cell function and potential therapeutic targets.
- Published
- 2021
19. Tumor cells dictate anti-tumor immune responses by altering pyruvate utilization and succinate signaling in CD8
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Ilaria Elia, Jared H. Rowe, Sheila Johnson, Shakchhi Joshi, Giulia Notarangelo, Kiran Kurmi, Sarah Weiss, Gordon J. Freeman, Arlene H. Sharpe, and Marcia C. Haigis
- Subjects
Physiology ,Neoplasms ,Pyruvic Acid ,Immunity ,Succinic Acid ,Tumor Microenvironment ,Humans ,Cell Biology ,Lactic Acid ,CD8-Positive T-Lymphocytes ,Molecular Biology ,Pyruvate Carboxylase - Abstract
The tumor microenvironment (TME) is a unique metabolic niche that can inhibit T cell metabolism and cytotoxicity. To dissect the metabolic interplay between tumors and T cells, we establish an in vitro system that recapitulates the metabolic niche of the TME and allows us to define cell-specific metabolism. We identify tumor-derived lactate as an inhibitor of CD8
- Published
- 2021
20. Tumor Cells Dictate Anti-Tumor Immune Responses by Altering Pyruvate Utilization and Succinate Signaling in Cd8 + T Cells
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Ilaria Elia, Giulia Notarangelo, Marcia C. Haigis, Sheila E. Johnson, Shakchhi Joshi, Gordon J. Freeman, Kiran Kurmi, Jared H. Rowe, and Arlene H. Sharpe
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Citric acid cycle ,Tumor microenvironment ,medicine.anatomical_structure ,Chemistry ,Cell ,medicine ,Cytotoxic T cell ,Metabolism ,Pyruvate dehydrogenase complex ,Autocrine signalling ,Cell biology ,Pyruvate carboxylase - Abstract
The tumor microenvironment (TME) is a unique metabolic niche that can inhibit CD8+ T cell metabolism and cytotoxic activity. It has been a challenge to dissect the metabolic interplay between tumors and T cells. Consequently, the mechanisms whereby the TME regulates CD8+ T cell function are not completely understood. Here we establish an in vitro system that recapitulates the metabolic niche of the TME and allows us to define cell-specific metabolism. We identify tumor-derived lactate as a direct inhibitor of CD8+ T cell cytotoxicity, revealing an unexpected metabolic shunt in the tricarboxylic acid (TCA) cycle in CD8+ T cells. Metabolically fit cytotoxic T cells shunt succinate out of the TCA cycle to promote autocrine signaling via the succinate receptor (SUCNR1), a G-protein coupled cell surface receptor. This leaves CD8+ T cells reliant on pyruvate carboxylase (PC) to replenish TCA cycle intermediates (anaplerosis). In contrast, lactate reduces PC-mediated anaplerosis, thus depleting the TCA cycle and reducing SUCNR1 signaling. Notably, inhibition of pyruvate dehydrogenase (PDH) is sufficient to override lactate-mediated reprogramming, and to restore PC activity, succinate secretion, and autocrine activation of SUCNR1 both in vitro and in vivo using syngeneic tumor models. Thus, dissecting the CD8+ T cell-specific metabolic adaptation to the TME unexpectedly revealed that a TCA cycle succinate shunt and plasticity in pyruvate flux is a crucial determinant of CD8+ T cell anti-tumor activity. Furthermore, we identify PDH as a potential drug target to allow CD8+ T cells to retain cytotoxicity and overcome a lactate-rich TME. These studies demonstrate that the TME remodels CD8+ T cell pyruvate metabolism, leading to a reduction in CD8+ T cell cytotoxicity and anti-tumor immunity.
- Published
- 2021
21. Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8
- Author
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Jefte M, Drijvers, Jacob E, Gillis, Tara, Muijlwijk, Thao H, Nguyen, Emily F, Gaudiano, Isaac S, Harris, Martin W, LaFleur, Alison E, Ringel, Cong-Hui, Yao, Kiran, Kurmi, Vikram R, Juneja, Justin D, Trombley, Marcia C, Haigis, and Arlene H, Sharpe
- Subjects
Antineoplastic Agents ,CD8-Positive T-Lymphocytes ,Endoplasmic Reticulum ,Article ,Mice, Inbred C57BL ,Mice ,Cell Line, Tumor ,Neoplasms ,Autophagy ,Tumor Cells, Cultured ,Animals ,Ferroptosis ,Humans ,Female - Abstract
Metabolic constraints in the tumor microenvironment constitute a barrier to effective anti-tumor immunity and similarities in the metabolic properties of T cells and cancer cells impede the specific therapeutic targeting of metabolism in either population. To identify distinct metabolic vulnerabilities of CD8(+) T cells and cancer cells, we developed a high-throughput in vitro pharmacologic screening platform and used it to measure the cell type–specific sensitivities of activated CD8(+) T cells and B16 melanoma cells to a wide array of metabolic perturbations during antigen-specific killing of cancer cells by CD8(+) T cells. We illustrated the applicability of this screening platform by showing that CD8(+) T cells were more sensitive to ferroptosis than B16 and MC38 cancer cells. Overexpression of ferroptosis suppressor protein 1 (FSP1) or cytosolic GPX4 yielded ferroptosis-resistant CD8(+) T cells without compromising their function, while genetic deletion of the ferroptosis sensitivity–promoting enzyme acyl-CoA synthetase long-chain family member 4 (ACSL4) protected CD8(+) T cells from ferroptosis, but impaired anti-tumor CD8(+) T cell responses. Our screen also revealed high T cell–specific vulnerabilities for compounds targeting NAD(+) metabolism or autophagy and ER stress pathways. We focused the current screening effort on metabolic agents. However, this in vitro screening platform may also be valuable for rapid testing of other types of compounds to identify regulators of anti-tumor CD8(+) T-cell function and potential therapeutic targets.
- Published
- 2020
22. Nitrogen Metabolism in Cancer and Immunity
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Kiran Kurmi and Marcia C. Haigis
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Anabolism ,Transamination ,Nitrogen ,Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Neoplasms ,Tumor Microenvironment ,Animals ,Humans ,Nitrogen cycle ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Tumor microenvironment ,Cell growth ,Immunity ,Cell Biology ,Amino acid ,Cell biology ,Pyrimidines ,chemistry ,Purines ,Cancer cell ,030217 neurology & neurosurgery - Abstract
As one of the fundamental requirements for cell growth and proliferation, nitrogen acquisition and utilization must be tightly regulated. Nitrogen can be generated from amino acids and utilized for biosynthetic processes through transamination and deamination reactions. Importantly, limitations of nitrogen availability in cells can disrupt the synthesis of proteins, nucleic acids and other important nitrogen-containing compounds. Rewiring cellular metabolism to support anabolic processes is a feature common to both cancer and proliferating immune cells. In this review, we will discuss how nitrogen is utilized in biosynthetic pathways and highlight different metabolic and oncogenic programs that alter the flow of nitrogen in order to sustain biomass production and growth - an important emerging feature of cancer and immune cell proliferation.
- Published
- 2019
23. Intercellular nanotubes mediate mitochondrial trafficking between cancer and immune cells
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Tanmoy, Saha, Chinmayee, Dash, Ruparoshni, Jayabalan, Sachin, Khiste, Arpita, Kulkarni, Kiran, Kurmi, Jayanta, Mondal, Pradip K, Majumder, Aditya, Bardia, Hae Lin, Jang, and Shiladitya, Sengupta
- Subjects
Mice, Inbred C57BL ,Mitochondrial Proteins ,Nanotubes ,Base Sequence ,Cell Line, Tumor ,Neoplasms ,Immunity ,Leukocytes ,Animals ,Humans ,Mitochondria - Abstract
Cancer progresses by evading the immune system. Elucidating diverse immune evasion strategies is a critical step in the search for next-generation immunotherapies for cancer. Here we report that cancer cells can hijack the mitochondria from immune cells via physical nanotubes. Mitochondria are essential for metabolism and activation of immune cells. By using field-emission scanning electron microscopy, fluorophore-tagged mitochondrial transfer tracing and metabolic quantification, we demonstrate that the nanotube-mediated transfer of mitochondria from immune cells to cancer cells metabolically empowers the cancer cells and depletes the immune cells. Inhibiting the nanotube assembly machinery significantly reduced mitochondrial transfer and prevented the depletion of immune cells. Combining a farnesyltransferase and geranylgeranyltransferase 1 inhibitor, namely, L-778123, which partially inhibited nanotube formation and mitochondrial transfer, with a programmed cell death protein 1 immune checkpoint inhibitor improved the antitumour outcomes in an aggressive immunocompetent breast cancer model. Nanotube-mediated mitochondrial hijacking can emerge as a novel target for developing next-generation immunotherapy agents for cancer.
- Published
- 2019
24. BRCA1 Deficiency Upregulates NNMT, Which Reprograms Metabolism and Sensitizes Ovarian Cancer Cells to Mitochondrial Metabolic Targeting Agents
- Author
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Taro Hitosugi, Catherine J. Huntoon, Daniel R. O'Brien, Cristina Correia, Xiaonan Hou, Scott H. Kaufmann, Hu Li, Arun Kanakkanthara, Thomas L. Ekstrom, Ethan P. Heinzen, Ann L. Oberg, S. John Weroha, Emma R. Purfeerst, Andrea E. Wahner Hendrickson, Kiran Kurmi, Sean C. Dowdy, and Larry M. Karnitz
- Subjects
0301 basic medicine ,Cancer Research ,endocrine system diseases ,DNA repair ,Carcinoma, Ovarian Epithelial ,Tigecycline ,Oxidative Phosphorylation ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Downregulation and upregulation ,Cell Line, Tumor ,Glucose import ,Antineoplastic Combined Chemotherapy Protocols ,Transcriptional regulation ,Hydroxybenzoates ,Nicotinamide N-Methyltransferase ,Animals ,Humans ,skin and connective tissue diseases ,Promoter Regions, Genetic ,Regulation of gene expression ,Ovarian Neoplasms ,Chemistry ,BRCA1 Protein ,Ovary ,Hydrazones ,DNA Methylation ,Triazoles ,Xenograft Model Antitumor Assays ,Cyclin-Dependent Kinases ,Mitochondria ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,DNA methylation ,Mutation ,Cancer research ,Female ,Homologous recombination ,Energy Metabolism ,Reprogramming - Abstract
BRCA1 plays a key role in homologous recombination (HR) DNA repair. Accordingly, changes that downregulate BRCA1, including BRCA1 mutations and reduced BRCA1 transcription, due to promoter hypermethylation or loss of the BRCA1 transcriptional regulator CDK12, disrupt HR in multiple cancers. In addition, BRCA1 has also been implicated in the regulation of metabolism. Here, we show that reducing BRCA1 expression, either by CDK12 or BRCA1 depletion, led to metabolic reprogramming of ovarian cancer cells, causing decreased mitochondrial respiration and reduced ATP levels. BRCA1 depletion drove this reprogramming by upregulating nicotinamide N-methyltransferase (NNMT). Notably, the metabolic alterations caused by BRCA1 depletion and NNMT upregulation sensitized ovarian cancer cells to agents that inhibit mitochondrial metabolism (VLX600 and tigecycline) and to agents that inhibit glucose import (WZB117). These observations suggest that inhibition of energy metabolism may be a potential strategy to selectively target BRCA1-deficient high-grade serous ovarian cancer, which is characterized by frequent BRCA1 loss and NNMT overexpression. Significance: Loss of BRCA1 reprograms metabolism, creating a therapeutically targetable vulnerability in ovarian cancer.
- Published
- 2019
25. Impaired β-cell glucokinase as an underlying mechanism in diet-induced diabetes
- Author
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Taro Hitosugi, Kiran Kurmi, Aleksey V. Matveyenko, Brian Lu, Egon J. Jacobus Ambuludi, Miguel Munoz-Gomez, Yogish C. Kudva, Yasuhiro Ikeda, Jason M. Tonne, and Kuntol Rakshit
- Subjects
Male ,0301 basic medicine ,Cell ,Intracellular Space ,lcsh:Medicine ,Medicine (miscellaneous) ,Impaired glucose tolerance ,Immunology and Microbiology (miscellaneous) ,Transduction, Genetic ,Insulin-Secreting Cells ,Glucokinase ,Insulin ,Glycolysis ,Islet biology ,geography.geographical_feature_category ,Insulin secretion ,Dependovirus ,Islet ,Potassium channel ,Up-Regulation ,medicine.anatomical_structure ,Signal Transduction ,Research Article ,lcsh:RB1-214 ,medicine.medical_specialty ,Neuroscience (miscellaneous) ,Carbohydrate metabolism ,Biology ,Diet, High-Fat ,General Biochemistry, Genetics and Molecular Biology ,Diabetes Mellitus, Experimental ,03 medical and health sciences ,Gene therapy ,Internal medicine ,Diabetes mellitus ,lcsh:Pathology ,medicine ,Animals ,Cell Proliferation ,geography ,Diet-induced diabetes ,lcsh:R ,Glucose Tolerance Test ,medicine.disease ,Mice, Inbred C57BL ,Glucose ,030104 developmental biology ,Endocrinology ,Calcium - Abstract
High-fat diet (HFD)-fed mouse models have been widely used to study early type 2 diabetes. Decreased β-cell glucokinase (GCK) expression has been observed in HFD-induced diabetes. However, owing to its crucial roles in glucose metabolism in the liver and in islet β-cells, the contribution of decreased GCK expression to the development of HFD-induced diabetes is unclear. Here, we employed a β-cell-targeted gene transfer vector and determined the impact of β-cell-specific increase in GCK expression on β-cell function and glucose handling in vitro and in vivo. Overexpression of GCK enhanced glycolytic flux, ATP-sensitive potassium channel activation and membrane depolarization, and increased proliferation in Min6 cells. β-cell-targeted GCK transduction did not change glucose handling in chow-fed C57BL/6 mice. Although adult mice fed a HFD showed reduced islet GCK expression, impaired glucose tolerance and decreased glucose-stimulated insulin secretion (GSIS), β-cell-targeted GCK transduction improved glucose tolerance and restored GSIS. Islet perifusion experiments verified restored GSIS in isolated HFD islets by GCK transduction. Thus, our data identify impaired β-cell GCK expression as an underlying mechanism for dysregulated β-cell function and glycemic control in HFD-induced diabetes. Our data also imply an etiological role of GCK in diet-induced diabetes. This article has an associated First Person interview with the first author of the paper., Summary: β-cell glucokinase expression is decreased in diet-induced diabetes. β-cell-targeted overexpression of glucokinase improved the diabetic phenotype, suggesting an etiological role of glucokinase downregulation in diet-induced diabetes.
- Published
- 2018
26. Circulating Mycobacterium bovis Peptides and Host Response Proteins as Biomarkers for Unambiguous Detection of Subclinical Infection
- Author
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Meetu Seth, My Yang, Lucy Vulchanova, W. Ray Waters, Tyler C. Thacker, Elise A. Lamont, Kiran Kurmi, Srinand Sreevatsan, Harish K Janagama, and Joao Ribeiro-Lima
- Subjects
Veterinary Medicine ,Microbiology (medical) ,Proteome ,Paratuberculosis ,Sensitivity and Specificity ,Microbiology ,Blood serum ,Bacterial Proteins ,Latent Tuberculosis ,medicine ,Animals ,Pathogen ,Subclinical infection ,Mycobacterium kansasii ,Mycobacterium bovis ,biology ,Clinical Laboratory Techniques ,Vitamin D-Binding Protein ,Mycobacteriology and Aerobic Actinomycetes ,biology.organism_classification ,medicine.disease ,Blood proteins ,Virology ,Cattle ,Peptides ,Tuberculosis, Bovine ,Biomarkers ,Blood Chemical Analysis ,Mycobacterium - Abstract
Bovine tuberculosis remains one of the most damaging diseases to agriculture, and there is also a concern for human spillover. A critical need exists for rapid, thorough, and inexpensive diagnostic methods capable of detecting and differentiating Mycobacterium bovis infection from other pathogenic and environmental mycobacteria at multiple surveillance levels. In a previous study, Seth et al. (PLoS One 4:e5478, 2009, doi:10.1371/journal.pone.0005478 ) identified 32 host peptides that specifically increased in the blood serum of M. bovis -infected animals). In the current study, 16 M. bovis proteins were discovered in the blood serum proteomics data sets. A large-scale validation analysis was undertaken for selected host and M. bovis proteins using a cattle serum repository containing M. bovis ( n = 128), Mycobacterium kansasii ( n = 10), and Mycobacterium avium subsp. paratuberculosis ( n = 10), cases exposed to M. bovis ( n = 424), and negative controls ( n = 38). Of the host biomarkers, vitamin D binding protein (VDBP) showed the greatest sensitivity and specificity for M. bovis detection. Circulating M. bovis proteins, specifically polyketide synthetase 5, detected M. bovis -infected cattle with little to no seroreactivity against M. kansasii - and M. avium subsp. paratuberculosis -infected animals. These data indicate that host and pathogen serum proteins can serve as reliable biomarkers for tracking M. bovis infection in animal populations.
- Published
- 2014
27. Tyrosine Phosphorylation of Mitochondrial Creatine Kinase 1 Enhances a Druggable Tumor Energy Shuttle Pathway
- Author
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Kiran Kurmi, Felix Boakye-Agyeman, Yuichi J. Machida, Taro Hitosugi, Matthew P. Goetz, Sadae Hitosugi, Scott H. Kaufmann, Thomas R. Larson, Judy C. Boughey, Elizabeth K. Wiese, Larry M. Karnitz, Liewei Wang, Qing Dai, Zhenkun Lou, and Jia Yu
- Subjects
0301 basic medicine ,Phosphocreatine ,Receptor, ErbB-2 ,Physiology ,Mice, Nude ,Antineoplastic Agents ,Breast Neoplasms ,Lapatinib ,Creatine ,Mitochondrial Proteins ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Animals ,Humans ,Phosphorylation ,Tyrosine ,skin and connective tissue diseases ,Creatine Kinase ,neoplasms ,Molecular Biology ,Kinase ,Tyrosine phosphorylation ,Cell Biology ,Trastuzumab ,Xenograft Model Antitumor Assays ,Mitochondria ,Cell biology ,Cell Transformation, Neoplastic ,030104 developmental biology ,Energy Transfer ,chemistry ,Drug Resistance, Neoplasm ,Creatinine ,Gene Knockdown Techniques ,Female ,Energy Metabolism ,Tyrosine kinase ,medicine.drug - Abstract
Summary How mitochondrial metabolism is altered by oncogenic tyrosine kinases to promote tumor growth is incompletely understood. Here, we show that oncogenic HER2 tyrosine kinase signaling induces phosphorylation of mitochondrial creatine kinase 1 (MtCK1) on tyrosine 153 (Y153) in an ABL-dependent manner in breast cancer cells. Y153 phosphorylation, which is commonly upregulated in HER2 + breast cancers, stabilizes MtCK1 to increase the phosphocreatine energy shuttle and promote proliferation. Inhibition of the phosphocreatine energy shuttle by MtCK1 knockdown or with the creatine analog cyclocreatine decreases proliferation of trastuzumab-sensitive and -resistant HER2 + cell lines in culture and in xenografts. Finally, we show that cyclocreatine in combination with the HER2 kinase inhibitor lapatinib reduces the growth of a trastuzumab-resistant HER2 + patient-derived xenograft. These findings suggest that activation of the phosphocreatine energy shuttle by MtCK1 Y153 phosphorylation creates a druggable metabolic vulnerability in cancer.
- Published
- 2018
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