TIPARP Overexpression: A Promising Strategy to Inhibit Oncogenic Phenotypes in TN-IBC
Discipline: Biological Sciences
Subcategory: cancer
Session: 3
Room: 1 - Hanover DE
Bibiana Torres-Colón - University of Puerto Rico, Río Piedras Campus
Co-Author(s): Ayeisha N. Colón-Ortiz, University of Puerto Rico-Río Piedras, PR; Keishla Rodríguez-Martir PhD, University of Puerto Rico-Río Piedras, PR; Esther A. Peterson-Peguero PhD, University of Puerto Rico-Río Piedras, PR
Triple-Negative Inflammatory Breast Cancer (TN-IBC) is a rare and aggressive subtype of breast cancer characterized by the absence of estrogen receptor alpha (ERα-66), progesterone receptor (PR), and HER2 amplification. TN-IBC is associated with rapid progression, poor prognosis, and limited treatment options, as therapies targeting alternative receptors, such as ERα-36 and GPR30, remain unexplored. Previous work in our lab showed that the phytoestrogen drug Coumestrol induces the upregulation of TIPARP, a novel tumor suppressor in TN-IBC. This study evaluates the effects of TIPARP overexpression on pro-oncogenic phenotypes and key signaling pathways in TN-IBC cell lines.
TIPARP was overexpressed in SUM149 cells through transfection with a customized TIPARP vector. Functional assays were performed to assess migration (wound healing assay), invasion (Boyden chamber assay), and apoptosis (Annexin V staining). Quantitative analyses of wound closure, cell invasion, and apoptotic markers were conducted using ImageJ and fluorescence microscopy. Expression levels of key oncogenic signaling molecules, AKT and ERK, were analyzed post-transfection through Western blotting.
TIPARP overexpression significantly inhibited cell migration (p < 0.0001) and induced early apoptosis (p < 0.0127), as evidenced by increased Annexin V staining compared to cells with no overexpresssion. Although TIPARP overexpression decreased cell invasion, it was not statistically significant. Western blot analysis confirmed that TIPARP overexpression led to decreased expression of AKT and ERK, two key regulators of pro-oncogenic signaling pathways. These results suggest the hypothesis that TIPARP acts as an inhibitor of pro-oncogenic phenotypes by indirectly regulating kinases AKT and ERK. These findings highlight TIPARP's potential to suppress oncogenic phenotypes by modulating critical pathways involved in cancer cell survival and motility while inducing apoptosis in TN-IBC cells. Future research will focus on elucidating the precise mechanistic interactions between TIPARP and the AKT and ERK pathways to better understand its tumor-suppressive role and potential as a therapeutic target.
References: Rodríguez-Mártir, K. M. (2022). Phytoestrogen Coumestrol as an anti-cancer therapy against triple-negative inflammatory breast cancer (thesis).
Cheng L, Li Z, Huang YZ, Zhang X, Dai XY, Shi L, Xi PW, Wei JF, Ding Q. TCDD-Inducible Poly-ADP-Ribose Polymerase (TIPARP), A Novel Therapeutic Target Of Breast Cancer. Cancer Manag Res. 2019;11:8991-9004 https://doi.org/10.2147/CMAR.S219289.
Funder Acknowledgement(s): I thank the IDGeNe Program, the NHGRI at the NIH, and specially, the members of the Peterson Breast Cancer Research Lab. Special thanks to Ayeisha N. Colon-Ortiz, and Dalissa Negrón-Figueroa for their unconditional mentorship and to Keishla Rodríguez-Martir for the RNAseq that originated this project. This project is funded by NIH grant 1R21CA253609-01 to Esther Peterson, PhD, Principal Investigator and Faculty Advisor at the University of Puerto Rico at Río Piedras, San Juan, Puerto Rico. This work was also funded by NIH grant 1R25HG012702-01 to José E. García-Arrarás, PhD, IDGeNe Research Program Director at the University of Puerto Rico at Río Piedras, San Juan, Puerto Rico.
Faculty Advisor: Esther A. Peterson-Peguero, PhD, esther.peterson@upr.edu
Role: I was responsible for conducting functional assays to overexpress TIPARP in TN-IBC cells, including wound healing, Boyden chamber, and Annexin V detection assays. I also performed Western blotting. These experiments evaluated TIPARP's effects on cell migration, invasion, apoptosis, and oncogenic signaling pathways. Additionally, I analyzed data and investigated changes in key regulators, such as AKT and ERK, to determine TIPARP's role in inhibiting pro-oncogenic phenotypes.