We have made significant progress in accomplishing the overall goals of our grant application. We have obtained skin cell from 14 subjects with various inherited gastrointestinal disorders, and we are in the process of reprogramming these cells to generate pluripotent stem cells. These cells will eventually be used to generate intestines and other tissue that will allow us to model the disorders of these various subjects. We anticipate that this approach will allow us to further develop tools that will let us understand the basis of chronic diarrhea and intestinal failure in children.
We have also performed total genome (exome) sequencing of several subjects with unique forms of chronic diarrhea and intestinal failure, and have identified several excellent candidate genes that likely account for these disorders. We will continue to screen other subjects with similar disorders to determine the full range of genes that are likely associated with these disorders.
We have also made considerable progress in our attempt to grow human small intestinal mucosa. Prior to our work, the small intestine had never been successfully grown and maintained in culture for more than just a few days. We utilized our experience working with murine intestine, and developed a novel method to grow human small intestines in a petri dish. We are now working to adapt our methods to grow intestinal samples that are obtained from endoscopic biopsies. We believe that this advancement will improve the usefulness of our culture system to samples obtained at the time of biopsy.
Taken together, we have made considerable progress in developing the tools required to understand the genetic basis of chronic diarrhea and intestinal failure in children. We are using a combination of genome sequencing, pluripotent stem cells, and tissue (somatic) stem cells to decipher the biology of established disorders, and to discover new disorders that have a significant impact on the growth and well being of children severe gastrointestinal disorders.