Abstract P2-06-06: Targeting loss of isoenzyme diversity as a novel therapeutic strategy in breast cancer
Authors: Marczyk M , Gunasekharan V , Wali VB , Shi W , Patwardhan G , Qing T , Pusztai L , Hatzis C 
Published: May 2019

Background: Several metabolic steps are mediated by distinct proteins or isoenzymes that catalyze the same reaction, providing redundancy of metabolic functions. Metabolic states are frequently altered in cancer to support survival and proliferation in hypoxic and otherwise hostile microenvironments, and metabolic re-wiring often involve loss of isoenzyme diversity. We hypothesize that targeting enzymes that have lost isoenzyme diversity in cancer, but not in normal cells, provides an opportunity to selectively target cancers. In this study, we assessed mRNA expression of all known human isoenzyme families in breast cancer and normal breast tissue and identified isoenzymes with loss of diversity within each breast cancer subtype.

Methods: We obtained RNAseq data from cancer and patient-matched normal breast tissues from the TCGA (N=66 HR+, N=24 HER2+, and N=15 TNBC tumors). We retrieved annotated human isoenzyme families from the ENZYME nomenclature database. We compared expression in cancer and matched normal samples from the same patient to identify isoenzymes that had i) same or increased expression of the target isoenzyme in cancer vs normal and ii) reduced expression of the complementary isoenzymes in cancer. We developed five scores that capture various elements of these characteristics and prioritized candidates as targets based on clustering and their combined ranking based on the five scores. We validated overexpression of the candidate isoenzymes relative to other isoforms in breast cancer microarray data from ArrayExpress (E-GEOD-76250: 33 TNBC, and E-GEOD-70951: 30 TNBC, 108 HR+, 10 HER2+).

Results: We identified 321 enzymes in the TCGA discovery cohort that correspond to 829 unique isoenzymes. Overall, 636, 483 and 429 isoenzymes were differentially expressed in HR+, HER2+ and TNBC cancers, respectively, compared to corresponding normal samples. Of these, 308 isoenzymes were differentially expressed relative to normal in all 3 subtypes. In all, 112 and 92, and 84 were selected as candidate isoenzyme therapeutic targets in HR+, HER2+ and TNBC, respectively. 23 isoenzymes prioritized in clustering step were further validated. Finally, 6 isoenzymes were validated in HR+ (ALDOA, GUSB, GYG1, MIF, P3H1, PCK2), 10 in HER2+ (ALDH1L2, ALDOA, GLYATL2, GUSB, GYG1, GYS1, MIF, P3H1, PCK2, PTGS1) and 12 in TNBC (ADSS, ALAS1, ALDH1L2, ALDOA, ART3, GLYATL2, GUSB, GYS1, HS3ST1, MIF, PCK2, SOAT1), as potential targets for breast cancer treatment. Of these, 5 potential isoenzyme targets (ALDH1L2, GUSB, GLYATL2, MIF, PCK2), which were mostly hydrolases and transferases, were further selected for ongoing experimental validation in the laboratory. Decreased expression of the complementary isoforms of these 5 targets were primarily due to DNA methylation of the genes in cancer.

Conclusions: We found that loss of isoenzyme diversity is a broad phenomenon in breast cancers that may be explored therapeutically. We identified several instances of “isoenzyme addiction” in which cancers depend exclusively on a single isoenzyme while downregulating via methylation the complementary isoenzymes, providing cancer-specific targeting opportunities. We are currently validating several of these targets in cell line models.

Citation Format: Marczyk M, Gunasekharan V, Wali VB, Shi W, Patwardhan G, Qing T, Pusztai L, Hatzis C. Targeting loss of isoenzyme diversity as a novel therapeutic strategy in breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-06.