Submission ID 93303
| Poster Code | HR-P-83 |
|---|---|
| Title of Abstract | ABROGATING TRIPLE NEGATIVE BREAST CANCER METASTASIS THROUGH INHIBITION OF CALPAIN-1/2 |
| Abstract Submission | Clinical literature shows that high expression of calpain-1/2 correlates with poor survival in breast cancer. Proteolytic cleavage of calpain-1/2 substrates in cancer is known to affect cell survival signaling, migration, invasion, and sensitivity to chemotherapeutics. Therefore, we hypothesized that genetic abrogation of calpain-1 and/or calpain-2 activity will make MDA-MB-231 human triple-negative breast cancer (TNBC) cells less motile, more sensitive to chemotherapeutic challenges, and will reduce their tumorigenic and metastatic potential. We developed a panel of CAPN1, CAPN2, and CAPNS1 genetic knock-out and lentiviral add-back MDA-MB-231 cell lines. We showed that we can successfully abrogate/restore calpain expression and activity. We demonstrate a phenotype of diminished cell migration in vitro, induced by calpain-1 deficiency through CAPN1 or CAPNS1 knock-out and rescued by the respective add-back. We also demonstrate reduced metastatic potential of these calpain-deficient TNBC cells in a mouse orthotopic engraftment model, while growth of a primary tumor is unaltered. To develop a pharmacological approach to induce the above-mentioned phenotypes, we developed a split NanoLuciferse reconstitution biosensor to measure the PEF-PEF interactions in calpain-1/2 and to screen for single amino acid substitutions that can disrupt the PEF-PEF dimerization that would be indicative of potential small-molecule binding sites. With the biosensor, we have demonstrated differences in PEF-PEF interactions of CAPN1/2 with CAPNS1, and we have employed these biosensors in an in vitro and an in silico high-throughput screening campaign with the eventual goal of developing allosteric isoform-specific inhibitors of calpain-1 and calpain-2 for in vivo applications. P.G. holds a Canadian Institutes of Health Research (CIHR) grant that supports the above-mentioned research, and I.Sh. is a recipient of The Canada Graduate Scholarships - Doctoral (CGS D). |
| Please indicate who nominated you | Queen's University, Dept Pathology and Molecular Medicine |
| What Canadian Institutes of Health Research (CIHR) institute is your research most closely aligned? | Cancer Research Genetics |
| What Canadian Institutes of Health Research (CIHR) pillar of health research does your research fall under? | Biomedical |
| PDF of abstract | ISh_CSHRF_2023_Abstract.pdf 2023-02-13 at 12:40:20 |
| Presenter and Author(s) | Ivan Shapovalov Ivan Shapovalov Yan Gao Kazem Nouri Peter Greer |
