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A Unique Opportunity to Test Whether Cell Fusion is a Mechanism of Breast Cancer Metastasis

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Annual rept. 1 Jul 2012-30 Jun 2013

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The goal of this proposal is to determine whether cell fusion between tumor cells and hematopoietic cells is the precipitating event for breast cancer metastasis and whether viral fusion proteins enable or catalyze this event. If successful, this discovery would dramatically change our approach to breast cancer therapy in the following specific ways. To date we have completed a significant portion of the tasks delineated in Aims 1 and 2. First we have optimized protocols for the separation of myeloid and monocyte populations from human mononuclear cell populations. Last cycle we optimized electroporation conditions for T47D and human mesenchymal stem cell populations and this cycle we have improved our transfection caliber by creating and optimizing lentiviral constructs for delivery of our fusion reporter BiFC pairs. As a result we have been able to complete our co-culture experiments to determine whether human breast cancer cells fuse spontaneously with hematopoietic cell types. Preliminary results suggest these populations do fuse spontaneously and that fusion products formed in this way can survive several days and are capable of proliferation. Unfortunately, for some cell types our original BiFC pairs did not allow proliferation and so this cycle we reengineered the pair to allow for proliferation of all cell types. Given this unanticipated problem, we have requested an extension and believe we can complete the aims of the grant in this period. In particular, we will purify spontaneous fusion products via flow cytometry and assess for the capacity to migrate and proliferate in vitro. Also, given our compelling early findings, we will investigate the frequency of spontaneous fusion in vivo and be able to discern whether fusion enables breast cancer metastasis.

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  • Biology
  • Medicine and Medical Research

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