TRANSDUCTION OF TAT/PTD ANTIAPOPTOTIC FUSION PROTEINS IN PANCREATIC ISLETS

INTRODUCTION. With the development of new strategies to avoid immunological rejection, transplantation of pancreatic islets has become a therapeutic reality to cure diabetes (1-2). However, despite the progress in islet isolation procedures, a single donor transplant does not provide enough islets to attain insulin independence. There is evidence that significant loss of islet cells takes place during isolation due to the triggering of apoptosis (3). There is also substantial evidence that reduction of isolation-induced apoptosis can improve the success rates of islet transplantation. The goal of this proposal is to address the needs for reduced apoptosis and improved viability of islets in conjunction with islet isolation procedures. We present in this study novel transduction methods that allow manipulation of islets to reduce apoptosis. Proteins can be directly transferred to cells when they are linked to protein transduction domains (PTDs), small peptide domains that can freely cross cell membranes. In particular, proteins fused to an 11-amino acid PTD from the human immunodeficiency virus, the TAT protein, readily diffuse across membranes and are efficiently transduced into virtually any cell type (4). The expression as well as the biological function of the TAT fusion protein are temporary without permanent modification of the cell sensitivity to apoptosis, thus avoiding undesirable long-term effects.