Huang-The goal is to investigate cancer specific nucleolar-genome interactions and their role in carcinogenesis. Specifically, we will work with perinucleolar compartment (PNC) that is physically associated with nucleoli and is nucleated upon multiple genome regions. Our previous findings indicate that PNC formation positively correlates with metastatic potential of cancer cells. Using PNC as a malignant cancer surrogate marker, we have developed a PNC inhibitor that shows selective anti-metastasis effect in vivo. We propose here to determine the functional interplay between nucleoli and PNCs in cancer cells. We will investigate whether PNCs mediate the association of cancer specific NADs to the nucleolus and influence their expression. We will evaluate the role the cancer specific PNC associated NADs in cancer development and metastasis. Foltz-The objective of the work is to investigate the patterns and mechanisms of genomic organization in the context to human disease. Specifically, the Foltz lab will investigate the associated between the centromere and the nucleolus in human primary cells and cell lines. These experiments are encompassed within Aim 3 of the U54 proposal. The experiments will include, but are not limited to, co-immunoprecipitation, LacO/LacI interaction assays, CRISPR/dCAS9 APEX labeling, immunofluorescence of endogenous proteins at the centromere and SNAP-Tag experiments. The Foltz lab will conduct these experiments in primary cell lines containing neocentromeres as well cultured cell lines. All experiments shall be completed in triplicate to ensure reproducibility of the results and will be complete in as timely fashion as possible. All experiments shall be carried out by experienced technical staff in the lab. Results and progress will be communicated between PI and co-investigators on at least a quarterly basis. The Foltz lab shall provide the results in the form of immunofluorescence data, immunoblots and protein lists from Mass Spectrometry. The Foltz lab shall provide individual raw data, as well as analyzed and summarized data. The Foltz shall provide publication quality figures for the dissemination of results in manuscript and presentation format.
|Effective start/end date||9/1/20 → 2/28/27|
- University of Massachusetts Medical School (OSP33101-00//5U01CA260699-02)
- National Cancer Institute (OSP33101-00//5U01CA260699-02)
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