title: Gene transfer of constitutively active Akt markedly improves human islet transplant outcomes in diabetic severe combined immunodeficient mice.
abstract: Akt is an important intracellular mediator of beta-cell growth and survival in rodents. However, whether constitutive activation of Akt in human beta-cells enhances the survival and function of transplanted islets is unknown. In the current study, we examined the efficacy of constitutive activation of Akt in improving human islet transplant outcomes using a marginal mass model in diabetic severe combined immunodeficient (SCID) mice. Human islets transduced with adenoviruses encoding constitutively active Akt1 (Adv-CA-Akt) displayed increased total and phosphorylated Akt and Akt kinase activity compared with control islets. Expression of CA-Akt in human islets induced a significant increase in beta-cell replication and a significant decrease in beta-cell death induced by serum and glucose deprivation or chronic hyperglycemia. Two control groups of islets (1,500 uninfected or adenovirus LacZ [Adv-LacZ]-transduced human islet equivalents [IEQs]) transplanted under the kidney capsule of streptozotocin-induced diabetic SCID mice were insufficient to correct hyperglycemia. Importantly and in marked contrast to these controls, 1,500 Adv-CA-Akt-transduced IEQs were capable of restoring euglycemia in diabetic SCID mice. Moreover, blood glucose normalization persisted for at least 6 months. Human plasma insulin at day 54 after transplant was 10-fold higher in Adv-CA-Akt islet recipients (2.4 +/- 0.4 ng/ml) compared with those receiving Adv-LacZ islets (0.25 +/- 0.08 ng/ml) (P < 0.05). In summary, expression of CA-Akt in human islets improves islet transplant outcomes in a subcapsular renal graft model in SCID mice. Akt is an attractive target for future strategies aimed at reducing the number of islets required for successful islet transplantation in humans.
authors.affiliation: Division of Endocrinology, BST-E-1140, University of Pittsburgh, 200 Lothrop St., Pittsburgh, PA 15261, USA.
authors.forename: Poornima
authors.initials: P
authors.lastname: Rao
authors.forename: Jennifer
authors.initials: J
authors.lastname: Roccisana
authors.forename: Karen K
authors.initials: KK
authors.lastname: Takane
authors.forename: Rita
authors.initials: R
authors.lastname: Bottino
authors.forename: Allan
authors.initials: A
authors.lastname: Zhao
authors.forename: Massimo
authors.initials: M
authors.lastname: Trucco
authors.forename: Adolfo
authors.initials: A
authors.lastname: García-Ocaña
mesh_terms: D000256:Adenoviridae; D000818:Animals; D016923:Cell Death; D049109:Cell Proliferation; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism* / Q000628:therapy; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D007515:Islets of Langerhans / Q000502:physiology; D016381:Islets of Langerhans Transplantation / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology*; D051057:Proto-Oncogene Proteins c-akt; D013997:Time Factors; D014183:Transplantation, Heterologous
chemical_list: D011518:Proto-Oncogene Proteins; C494918:AKT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt