As a faculty member in the Department of Radiation Oncology, I am actively involved in performing research in radiation biology. I actively participate in the Center for Medical Counter Measures Against Radiation (CMCR) grant awarded to the University of Pittsburgh as a core leader for the Radiobiological Standardization Core and an investigator in Project 1. I also organize and lecture in the radiation biology class for the radiation oncology residents. I also participate in instructing medical students from the University of Pittsburgh Medical School in how to perform radiation biology research. My research involves developing new radiation protectors and mitigators for protection against irradiation-induced damage in vitro and in vivo as well as studying the late effects of irradiation, effects of irradiation on pregnant mice, and development of ALS.
I am the coordinator and main lecturer for the radiation biology class for the radiation oncology residents. I also work closely with the radiation oncology residents during their research year. I also instruct medical students from the University of Pittsburgh Medical School as well as other medical schools who spend anywhere from one month to a year doing research in the Radiation Oncology Research Laboratory. This involves writing research proposals, abstracts and manuscripts, good laboratory technique, planning experiments, collecting data, and statistical analysis.
Sharlow Er, Leimgruber S, Lira A, McConnell MJ, Norambuena A, Bloom GS, Epperly MW, Greenberger JS, and Lazo JS. A small molecule screen exposes mTOR signaling pathway involvement in radiation-induced apoptosis, ACS Chem Biol. 11(5):1428-1437, 2016, PMID 26938669.
Huang Z, Epperly M, Watkins SC, Greenberger JS, Kagan VE, and Bayir H. Necrostatin-1 rescues mice from lethal irradiation. Biochim Biophys Acta, 1862(4):850-856. Doi:10.106/j.bbadis.2016.01.014. Epub 2016 Jan 20, 2016.
Shinde A, Berhane H, Rhieu BH, Kalash R, Wu K, Goff J, Epperly MW, Franicola D, Zhang X, Dixon T, Shields D, Wang H, Wipf P, Parmar K, Guinan E, Kagan V, Tyurin V, Ferris RL, Zhang, X, Li S, and Greenberger JS. Intraoral mitochondrial-targeted GS-nitroxide, JP4-039, radioprotects normal tissue in tumor-bearing radiosensitive Fancd2 (-/-) (C57BL/6) mice. Radiat Res, 185 (2):134-150, doi:10.1667/RR14035.1.Epub 2016 Jan 20, 2016.
Greenberger J, Kagan V, Bayir H, Wipf, and Epperly MW. Antioxidant approaches to management of ionizing irradiation injury. Antioxidants. Jan 23; 4 (1): 82-101. Doi: 10.3390/antiox4010082, 2015.
Gomez-Casal R, Epperly MW, Wang H, Proia DA, Greenberger JS and Levina V. Radioresistant human lung adenocarcinoma cells that survived multiple fractions of ionizing radiation are sensitive to HSP 90 inhibition. Oncotarget 6 (42):44306-44322.doi:10.18632/oncotarget.6248, 2015.
Wang Y, Li W, Patel SS, Cong J, Zhang N, Sabbatino F, Liu X, Qi Y, Huang P, Lee H, Taghian A, Li JJ, DeLeo AB, Ferrone S, Epperly MW, Ferrone CR, Ly A, Brachtel EF, Wang X. Blocking the formation of radiation-induced breast cancer stem cells. Oncotarget. 5(11):3743-55, 2015.
Gomez-Casal R, Bhattacharya C, Epperly MW, Basse PH, Wang H, Wang X, Proia DA, Greenberger JS, Socinski MA, Levina V. The HSP90 Inhibitor Ganetespib Radiosensitizes Human Lung Adenocarcinoma Cells. Cancers (Basel). 2015 May 22;7(2):876-907. PubMed PMID: 26010604.
Oczypok EA, Milutinovic PS, Alcorn JF, Khare A, Crum LT, Manni ML, Epperly MW, Pawluk AM, Ray A, Oury TD. Pulmonary receptor for advanced glycation end-products promotes asthma pathogenesis through IL-33 and accumulation of group 2 innate lymphoid cells. J Allergy Clin Immunol. 2015 Apr 27. pii: S0091-6749(15)00409-1. PubMed PMID: 25930197.
Wang Y, Li W, Patel SS, Cong J, Zhang N, Sabbatino F, Liu X, Qi Y, Huang P, Lee H, Taghian A, Li JJ, DeLeo AB, Ferrone S, Epperly MW, Ferrone CR, Ly A, Brachtel EF, and Wang X. Blocking the formation of radiation-induced breast cancer stem cells. Oncotarget 2015 5(11):3743, 3755. PubMed PMID: 25003837.
Glowacki J, Mizuno S, Kung J, Goff J, Epperly M, Dixon T, Wang H, and Greenberger JS. Effects of mouse genotype on bone wound healing and irradiation-induced delay of healing. 2014, In Vivo 28(2): 189-196. PubMed PMID: 24632972.
Rhieu BH, Shinde A, Epperly MW, Dixon T, Wang H, Chaillet R, Greenberger JS. Organ-specific responses of total body irradiated doxycycline-inducible manganese superoxide dismutase Tet/Tet mice. In Vivo. 2014 Nov-Dec;28(6):1033-43. PubMed PMID: 25398796.
Shinde A, Epperly MW, Cao S, Holt D, Goff J, Shields D, Franicola D, Wipf P, Wang H, Greenberger JS. Improved hematopoiesis in GS-nitroxide (JP4-039)-treated mouse long-term bone marrow cultures and radioresistance of derived bone marrow stromal cell lines. In Vivo. 2014 Sep-Oct;28(5):699-708. PubMed PMID: 25189880.
Shinde A, Epperly MW, Cao S, Franicola D, Shields D, Wang H, Wipf P, Sprachman MM, Greenberger JS. Effects of the bifunctional sulfoxide MMS350, a radiation mitigator, on hematopoiesis in long-term bone marrow cultures and on radioresistance of marrow stromal cell lines. In Vivo. 2014 Jul-Aug;28(4):457-65. PubMed PMID: 24982210.
Rhieu BH, Epperly MW, Cao S, Franicola D, Shields D, Goff J, Wang H, Greenberger JS. Increased hematopoiesis in long-term bone marrow cultures and reduced irradiation-induced pulmonary fibrosis in Von Willebrand factor homologous deletion recombinant mice. In Vivo. 2014 Jul-Aug;28(4):449-56. PubMed PMID: 24982209.
Rhieu BH, Epperly MW, Cao S, Goff J, Shields D, Franicola D, Wang H, Greenberger JS. Improved longevity of hematopoiesis in long-term bone marrow cultures and reduced irradiation-induced pulmonary fibrosis in Toll-like receptor-4 deletion recombinant-negative mice. In Vivo. 2014 Jul-Aug;28(4):441-8. PubMed PMID: 24982208.
Epperly MW, Goff JP, Franicola D, Wang H, Wipf P, Li S, Greenberger JS. Esophageal radioprotection by swallowed JP4-039/F15 in thoracic-irradiated mice with transgenic lung tumors. In Vivo. 2014 Jul-Aug;28(4):435-40. PubMed PMID: 24982207.
Berhane H, Shinde A, Kalash R, Xu K, Epperly MW, Goff J, Franicola D, Zhang X, Dixon T, Shields D, Wang H, Wipf P, Li S, Gao X, Greenberger JS. Amelioration of radiation-induced oral cavity mucositis and distant bone marrow suppression in fanconi anemia Fancd2-/- (FVB/N) mice by intraoral GS-nitroxide JP4-039. Radiat Res. 2014 Jul;182(1):35-49. doi: 10.1667/RR13633.1. Epub 2014 Jun 16. PubMed PMID: 24932534; PubMed Central PMCID: PMC4101533.
Tyurina YY, Poloyac SM, Tyurin VA, Kapralov AA, Jiang J, Anthonymuthu TS, Kapralova VI, Vikulina AS, Jung MY, Epperly MW, Mohammadyani D, Klein-Seetharaman J, Jackson TC, Kochanek PM, Pitt BR, Greenberger JS, Vladimirov YA, Bayır H, Kagan VE. A mitochondrial pathway for biosynthesis of lipid mediators. Nat Chem. 2014 Jun;6(6):542-52. PubMed PMID: 24848241; PubMed Central PMCID: PMC4201180.
Kalash R, Berhane H, Au J, Rhieu BH, Epperly MW, Goff J, Dixon T, Wang H, Zhang X, Franicola D, Shinde A, Greenberger JS. Differences in irradiated lung gene transcription between fibrosis-prone C57BL/6NHsd and fibrosis-resistant C3H/HeNHsd mice. In Vivo. 2014 Mar-Apr;28(2):147-71. PubMed PMID: 24632969; PubMed Central PMCID: PMC4074886.
Greenberger JS, Berhane H, Shinde A, Rhieu BH, Bernard M, Wipf P, Skoda EM, Epperly MW. Can Radiosensitivity Associated with Defects in DNA Repair be Overcome by Mitochondrial-Targeted Antioxidant Radioprotectors. Front Oncol. 2014 Feb 17;4:24. PubMed PMID: 24596683; PubMed Central PMCID: PMC3926189.
Kalash R, Berhane H, Yang Y, Epperly MW, Wang H, Dixon T, Rhieu B, Greenberger JS, Huq MS. Improved survival of mice after total body irradiation with 10 MV photon, 2400 MU/min SRS beam. In Vivo. 2014 Jan-Feb;28(1):1-12. PubMed PMID: 24425830; PubMed Central PMCID: PMC4046118.
Kanter DJ, O'Brien MB, Shi XH, Chu T, Mishima T, Beriwal S, Epperly MW, Wipf P, Greenberger JS, Sadovsky Y. The impact of ionizing radiation on placental trophoblasts. Placenta. 2014 Feb;35(2):85-91. PubMed PMID: 24418702; PubMed Central PMCID: PMC3954710.
Berhane H, Epperly MW, Goff J, Kalash R, Cao S, Franicola D, Zhang X, Shields D, Houghton F, Wang H, Wipf P, Parmar K, Greenberger JS. Radiologic differences between bone marrow stromal and hematopoietic progenitor cell lines from Fanconi Anemia (Fancd2(-/-)) mice. Radiat Res. 2014 Jan;181(1):76-89. PubMed PMID: 24397476; PubMed Central PMCID: PMC3970166.
Marrow from a Second strain of Double Knockout (DKO) SMAD3-/- Fancd2-/- Mice (Uniform 129/Sv background) Shows Marked Reduction of Duration of Hematopoiesis in Continuous Bone Marrow Cultures
Introduction: We previously reported that abrogation of TGF-β signaling in (DKO) SMAD3-/- (129/Sv) Fancd2-/- (B6) mice showed resistance of hematopoietic progenitors to inhibition of colony formation by TGF-β, but did not alter radiosensitivity of bone marrow stromal cells, or reduced longevity of hematopoiesis in vitro. SMAD3-/- (129/Sv) mice and derived cell lines were radioresistant while Fancd2-/- mice and bone marrow stromal cells were radiosensitive, consistent with the phenotype of the mice. Because studies of hematopoietic stem cell numbers in DKO mice require three control strains to account for the different genetic backgrounds of each of the single knockout mice (B6, 129/Sv, and B6 X 129/Sv F1), we derived a second DKO mouse strain in which both genes were inactivated in a single mouse strain (129/Sv). We sought to determine the biological properties of this second DKO strain.
Materials and Methods: 129/Sv control, as well as single and double knockout mouse strains on the 129/Sv background were sacrificed, and long-term bone marrow cultures were established according to published methods. Sensitivity of freshly removed bone marrow to TGF-β inhibition of colony formation (CFU-GEMM) was carried out with each mouse strain and results compared.
Results: DKO pups of this strain in which both genes were inactivated on the 129/Sv background (DKO #2) demonstrated growth and development similar to those of the first DKO strain (DKO #1). Fresh marrow CFU-GEMM colony forming progenitors from both SMAD3-/- and DKO mice were resistant to inhibition of CFU-GEMM colony formation in vitro by TGF-β.
Long-term bone marrow cultures from DKO #2 displayed reduced duration of hematopoiesis similar to those properties of DKO #1 mice. Cobblestone island formation (indicative of adherent hematopoietic cell colonies) was markedly reduced in both Fancd2-/- and DKO #2 mouse strains. SMAD3-/- marrow cultures showed increased duration of adherent hematopoietic cobblestone island forming cells similar to that found with DKO #1 mouse experiments. Non-adherent cell production was depressed in DKO (#2), as well as Fancd2-/- (129/Sv) long-term bone marrow cultures confirming results with DKO (#1) marrow. In contrast, DKO #2 long-term marrow cultures showed a more profound decrease in the generation of colony forming progenitor cells forming day 7 and day 14 colonies in vitro.
Therefore, inactivation of both SMAD3 and Fancd2 in the same genetic background mouse strain (DKO #2) produced the phenotype seen in DKO #1 and was consistent with the Fancd2-/- genotype, but retained resistance to TGF-β consistent with the SMAD3-/- genotype.
Conclusions: Abrogation of TGF-β signaling in a second Fancd2-/- (129/Sv) background mouse strain demonstrated abrogation of TGF-β mediated suppression of hematopoiesis. However, the dominant phenotype of DKO #2 mice was consistent with the Fancd2-/- genotype. Thus, in two different DKO mouse strains, a predominant phenotype was that of the Fancd2-/- mouse.
Faculty Support on Research grants is 65 percent
Source/Grant Number: NIAID/U19 AI06680201-11
Grant Title: Signature-Directed, Sequential Delivery of Radiation Mitigators
C: Radiobiological Standardization Core
Role in Project & Percentage of Effort: Director 3 calendar months
Years Inclusive: 9/01/15 to 8/31/20
Direct Dollars (total/annual): $135,561.04
Indirect Dollars (total/annual): $73,203.00
Source/Grant Number: NIAID/U19 AI06680201-11
Grant Title: Signature-Directed Sequential Delivery of Radiation Mitigators
Project 1: Signature-Directed Combination Mitigator Therapy Based on GS-nitroxides
Role in Project & Percentage of Effort: Co-Investigator 4 calendar months
Years Inclusive: 9/01/15 to 8/31/20
Direct Dollars (total/annual): $65,329.87
Indirect Dollars (total/annual): $35,278.13
Source/Grant Number: NIH/RO1 DKO71085-09
Grant Title: Roles of Nitric Oxide as Superoxide in Cystitis
Role in Project & Percentage of Effort: Co-Investigator 0.86 calendar months
Years Inclusive: 4/01/13 to 3/31/18
Direct Dollars (total/annual): $19,047
Indirect Dollars (total/annual): $9,904
Source/Grant Number: NIH/RO1 GM102989
Grant Title: Core C: Rational Design of Lipidic Vectors for Mitochondria-Targeted Antioxidants
Role in Project & Percentage of Effort: Co-Investigator 0.96 calendar months
Years Inclusive: 7/01/13 to 4/30/17
Direct Dollars (total/annual): $19,099
Indirect Dollars (total/annual): $9,931