Daniel A Pomeranz Krummel1, Tahseen H Nasti2, Milota Kaluzova3, Laura Kallay1, Debanjan Bhattacharya1, Johannes C Melms4, Benjamin Izar4, Maxwell Xu5, Andre Burnham3, Taukir Ahmed6, Guanguan Li6, David Lawson7, Jeanne Kowalski8, Yichun Cao9, Jeffrey M Switchenko10, Dan Ionascu11, James M Cook6, Mario Medvedovic12, Andrew Jenkins13, Mohammad K Khan14, Soma Sengupta15. 1. Department of Neurology and Rehabilitation Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio. 2. Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia. 3. Emory University School of Medicine, Atlanta, Georgia. 4. Columbia Center for Translational Immunology, Columbia University College of Physicians and Surgeons, New York, New York. 5. Johns Hopkins University, Baltimore, Maryland. 6. Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin. 7. Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory University, Atlanta, Georgia. 8. Department of Oncology, LIVESTRONG Cancer Institutes, Dell Medical School, University of Texas, Austin, Texas. 9. Biostatistics Shared Resource, Winship Cancer Institute of Emory University, Atlanta, Georgia. 10. Biostatistics Shared Resource, Winship Cancer Institute of Emory University, Atlanta, Georgia; Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, Georgia. 11. Department of Radiation Oncology, University of Cincinnati College of Medicine, Cincinnati, Ohio. 12. Department of Environmental Health, University of Cincinnati College of Medicine, Cincinnati, Ohio. 13. Departments of Anesthesiology, Pharmacology and Chemical Biology, Emory University School of Medicine, Atlanta, Georgia. 14. Department of Radiation Oncology, Winship Cancer Institute of Emory University, Atlanta, Georgia. 15. Department of Neurology and Rehabilitation Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio. Electronic address: sengupsm@ucmail.uc.edu.
Abstract
PURPOSE: Most patients with metastatic melanoma show variable responses to radiation therapy and do not benefit from immune checkpoint inhibitors. Improved strategies for combination therapy that leverage potential benefits from radiation therapy and immune checkpoint inhibitors are critical. METHODS AND MATERIALS: We analyzed metastatic melanoma tumors in the TCGA cohort for expression of genes coding for subunits of type A γ-aminobutyric acid (GABA) receptor (GABAAR), a chloride ion channel and major inhibitory neurotransmitter receptor. Electrophysiology was used to determine whether melanoma cells possess intrinsic GABAAR activity. Melanoma cell viability studies were conducted to test whether enhancing GABAAR mediated chloride transport using benzodiazepine-impaired viability. A syngeneic melanoma mouse model was used to assay the effect of benzodiazepine on tumor volume and its ability to potentiate radiation therapy or immunotherapy. Treated tumors were analyzed for changes in gene expression by RNA sequencing and presence of tumor-infiltrating lymphocytes by flow cytometry. RESULTS: Genes coding for subunits of GABAARs express functional GABAARs in melanoma cells. By enhancing GABAAR-mediated anion transport, benzodiazepines depolarize melanoma cells and impair their viability. In vivo, benzodiazepine alone reduces tumor growth and potentiates radiation therapy and α-PD-L1 antitumor activity. The combination of benzodiazepine, radiation therapy, and α-PD-L1 results in near complete regression of treated tumors and a potent abscopal effect, mediated by increased infiltration of polyfunctional CD8+ T cells. Treated tumors show expression of cytokine-cytokine receptor interactions and overrepresentation of p53 signaling. CONCLUSIONS: This study identifies an antitumor strategy combining radiation and/or an immune checkpoint inhibitor with modulation of GABAARs in melanoma using benzodiazepine.
PURPOSE: Most patients with metastatic melanoma show variable responses to radiation therapy and do not benefit from immune checkpoint inhibitors. Improved strategies for combination therapy that leverage potential benefits from radiation therapy and immune checkpoint inhibitors are critical. METHODS AND MATERIALS: We analyzed metastatic melanoma tumors in the TCGA cohort for expression of genes coding for subunits of type A γ-aminobutyric acid (GABA) receptor (GABAAR), a chloride ion channel and major inhibitory neurotransmitter receptor. Electrophysiology was used to determine whether melanoma cells possess intrinsic GABAAR activity. Melanoma cell viability studies were conducted to test whether enhancing GABAAR mediated chloride transport using benzodiazepine-impaired viability. A syngeneic melanomamouse model was used to assay the effect of benzodiazepine on tumor volume and its ability to potentiate radiation therapy or immunotherapy. Treated tumors were analyzed for changes in gene expression by RNA sequencing and presence of tumor-infiltrating lymphocytes by flow cytometry. RESULTS: Genes coding for subunits of GABAARs express functional GABAARs in melanoma cells. By enhancing GABAAR-mediated anion transport, benzodiazepines depolarize melanoma cells and impair their viability. In vivo, benzodiazepine alone reduces tumor growth and potentiates radiation therapy and α-PD-L1 antitumor activity. The combination of benzodiazepine, radiation therapy, and α-PD-L1 results in near complete regression of treated tumors and a potent abscopal effect, mediated by increased infiltration of polyfunctional CD8+ T cells. Treated tumors show expression of cytokine-cytokine receptor interactions and overrepresentation of p53 signaling. CONCLUSIONS: This study identifies an antitumor strategy combining radiation and/or an immune checkpoint inhibitor with modulation of GABAARs in melanoma using benzodiazepine.
Authors: H Friedman; D J Greenblatt; G R Peters; C M Metzler; M D Charlton; J S Harmatz; E J Antal; E C Sanborn; S F Francom Journal: Clin Pharmacol Ther Date: 1992-08 Impact factor: 6.875
Authors: Aravind Subramanian; Pablo Tamayo; Vamsi K Mootha; Sayan Mukherjee; Benjamin L Ebert; Michael A Gillette; Amanda Paulovich; Scott L Pomeroy; Todd R Golub; Eric S Lander; Jill P Mesirov Journal: Proc Natl Acad Sci U S A Date: 2005-09-30 Impact factor: 11.205
Authors: Jeffrey A Sosman; Kevin B Kim; Lynn Schuchter; Rene Gonzalez; Anna C Pavlick; Jeffrey S Weber; Grant A McArthur; Thomas E Hutson; Stergios J Moschos; Keith T Flaherty; Peter Hersey; Richard Kefford; Donald Lawrence; Igor Puzanov; Karl D Lewis; Ravi K Amaravadi; Bartosz Chmielowski; H Jeffrey Lawrence; Yu Shyr; Fei Ye; Jiang Li; Keith B Nolop; Richard J Lee; Andrew K Joe; Antoni Ribas Journal: N Engl J Med Date: 2012-02-23 Impact factor: 91.245
Authors: Andrew I Su; Tim Wiltshire; Serge Batalov; Hilmar Lapp; Keith A Ching; David Block; Jie Zhang; Richard Soden; Mimi Hayakawa; Gabriel Kreiman; Michael P Cooke; John R Walker; John B Hogenesch Journal: Proc Natl Acad Sci U S A Date: 2004-04-09 Impact factor: 11.205
Authors: S Turajlic; S J Furney; G Stamp; S Rana; G Ricken; Y Oduko; G Saturno; C Springer; A Hayes; M Gore; J Larkin; R Marais Journal: Ann Oncol Date: 2014-02-06 Impact factor: 32.976
Authors: Laura Kallay; Havva Keskin; Alexandra Ross; Manali Rupji; Olivia A Moody; Xin Wang; Guanguan Li; Taukir Ahmed; Farjana Rashid; Michael Rajesh Stephen; Kirsten A Cottrill; T Austin Nuckols; Maxwell Xu; Deborah E Martinson; Frank Tranghese; Yanxin Pei; James M Cook; Jeanne Kowalski; Michael D Taylor; Andrew Jenkins; Daniel A Pomeranz Krummel; Soma Sengupta Journal: J Neurooncol Date: 2019-02-06 Impact factor: 4.130
Authors: Debanjan Bhattacharya; Vaibhavkumar S Gawali; Laura Kallay; Donatien K Toukam; Abigail Koehler; Peter Stambrook; Daniel Pomeranz Krummel; Soma Sengupta Journal: Exp Biol Med (Maywood) Date: 2021-10