(830) Functional Analysis of GAPDHP65 Suggests a Role in Triple-Negative Breast Cancer Viability

Introduction/Rationale: Triple-negative breast cancer (TNBC) is an aggressive subtype with limited treatment options. Dysregulated pseudogene expression is increasingly recognized in cancer progression, though many remain poorly characterized. This study examines GAPDHP65, a pseudogene involved in TNBC growth and survival.

Methods: Computational analysis of gene expression profiles identified GAPDHP65 as highly expressed in TNBC compared to normal breast tissue. Experimental validation was conducted using the HCC-1937 TNBC cell line, where GAPDHP65 knockout was generated through CRISPR-Cas9 and editing efficiency was confirmed by indel detection and ligation sequencing. Cell viability and proliferation were measured in wild-type and knockout HCC-1937 cells using standard assays, forming the basis of our previously presented work. Current experiments are focused on expanding validation to the HTB-26 TNBC cell line and conducting detailed apoptosis and cell cycle analyses to further define GAPDHP65 function.

Results: GAPDHP65 expression was elevated in TNBC cells, and its knockout (validated by sequencing) reduced proliferation in HCC-1937 cells, suggesting a role in maintaining cell survival. Preliminary observations indicate similar trends in the HTB-26 line. Ongoing cell cycle and apoptosis analyses are aimed at determining whether GAPDHP65 loss alters S-phase progression or increases programmed cell death, which will further clarify its contribution to TNBC growth dynamics.

Conclusion: Our previous work demonstrated that GAPDHP65 knockout impairs TNBC cell survival and progression, revealing its potential as a therapeutic target. These findings suggested that GAPDHP65 supports tumor growth by influencing apoptosis and cell cycle progression. Building on this foundation, the current study expands the analysis to additional TNBC lines and investigates downstream molecular changes following GAPDHP65 loss.