Role of Mitochondrial Inheritance on Prostate Cancer Outcome in African American Men
Technical Report,30 Sep 2011,29 Sep 2015
Baylor College of Medicine Houston United States
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We are examining the hypothesis that mitochondrial inheritance plays a significant role in aggressiveness of prostate cancer in African Americans. In the first year of the project we identified 2,000 noncancerous tissues samples from African American men with prostate cancer and we have extracted DNA from 1,500 of them. We have validated a robust new DNA sequencing technique developed by our collaborator using single amplicon long-range PCR that permits deep coverage 10,000-20,000X on average of the mitochondrial genome. We have sequenced 652 samples derived from frozen fully using this technology. The additional DNA samples derived from paraffin embedded FFPE tissue were more challenging, but have now been sequenced. Mapping of DNA variants in our sequenced genomes to mitochondrial genes is nearly complete and examples of the data we have obtained are presented here. After months of testing, we determined that the ethidium bromide methodology for generating prostate cancer cell line cybrids was not effective and we have instead used a Rhodamine-6-G procedure. PNT1A cybrid cell lines have been generated and we have characterized these cells. PC3 cells have been more difficult, but we continue to work on this cell line. We anticipate the first manuscripts relating to the large amount of data generated in this project will be written and submitted in the spring of 2016. To our knowledge this represents the largest collection of mitochondrial sequence data from African Americans in existence.