P-200

Inhibition of tissue factor prolongs intrahepatic islet xenograft survival in diabetic SCID mouse model

Q. Liang¹, X. Ma¹, B. Ye¹, Q. Dong¹, S. Yi², J. Zhu¹, W. Wang^1
1 The Third Xiangya Hospital at Central South University, Cell Transplantation and Gene Therapy Institute, Changsha, China; ² Centre for Transplantation and Renal Research, Westmead Millennium Institute, Westmead Hospital, Sydney, Australia

Background: IBMIR is one of barriers to be overcome in islets transplantation. In this study, we investigated the effect of TF knockdown by antisense RNA transfection to block IBMIR in pig islet xenotransplantation with diabetic SCID mouse model.
Methods: Antisense RNAs were designed to transfect porcine islet cell with antisense technology, then real-time PCR was applied to select the best antisense RNA for TF silencing and the results were confirmed by western blot and FACS; to known the level of development of IBMIR, the indexes were detected in vitro model of IBMIR in the porcine islet cell after TF silencing. Further the viability of NICC in different treatment groups were examined by FACS. Glucose challenge test was also demonstrated to assess the toxicity. Each of eleven pairs of SCID mice (SCID) with streptozotocin-induced diabetes was transplanted with 500-1,000 NICC via portal vein. Experimental animals received NICC treated with TF antisense RNA, control mice received naïve NICC. Markers of coagulation and fasting blood-glucose were assessed post-Tx, higher fasting C-peptid(CP) level was assessed at end of study, 6 months post-Tx.
Results: TF expression of NICC was significantly down-regulated by the antisense RNA both at the RNA level and protein level. In a tubing loop model, NICC transfected with the TF-targeted antisense RNA showed an inhibition of IBMIR with an inhibition of coagulation and complement activation and less consumption of platelets and other blood cells. Compared to the control animals, experimental SCID had shown low posttransplant markers of coagulation, higher fasting CP levels (end of study) and prolonged graft function.
Conclusions: These data suggest that the transfection of islets with the TF-targeted antisense RNA can limit the effects of IBMIR, thereby prolong NICC graft survival in SCID model.

P-200

Inhibition of tissue factor prolongs intrahepatic islet xenograft survival in diabetic SCID mouse model

Q. Liang¹, X. Ma¹, B. Ye¹, Q. Dong¹, S. Yi², J. Zhu¹, W. Wang^1
1 The Third Xiangya Hospital at Central South University, Cell Transplantation and Gene Therapy Institute, Changsha, China; ² Centre for Transplantation and Renal Research, Westmead Millennium Institute, Westmead Hospital, Sydney, Australia

Background: IBMIR is one of barriers to be overcome in islets transplantation. In this study, we investigated the effect of TF knockdown by antisense RNA transfection to block IBMIR in pig islet xenotransplantation with diabetic SCID mouse model.
Methods: Antisense RNAs were designed to transfect porcine islet cell with antisense technology, then real-time PCR was applied to select the best antisense RNA for TF silencing and the results were confirmed by western blot and FACS; to known the level of development of IBMIR, the indexes were detected in vitro model of IBMIR in the porcine islet cell after TF silencing. Further the viability of NICC in different treatment groups were examined by FACS. Glucose challenge test was also demonstrated to assess the toxicity. Each of eleven pairs of SCID mice (SCID) with streptozotocin-induced diabetes was transplanted with 500-1,000 NICC via portal vein. Experimental animals received NICC treated with TF antisense RNA, control mice received naïve NICC. Markers of coagulation and fasting blood-glucose were assessed post-Tx, higher fasting C-peptid(CP) level was assessed at end of study, 6 months post-Tx.
Results: TF expression of NICC was significantly down-regulated by the antisense RNA both at the RNA level and protein level. In a tubing loop model, NICC transfected with the TF-targeted antisense RNA showed an inhibition of IBMIR with an inhibition of coagulation and complement activation and less consumption of platelets and other blood cells. Compared to the control animals, experimental SCID had shown low posttransplant markers of coagulation, higher fasting CP levels (end of study) and prolonged graft function.
Conclusions: These data suggest that the transfection of islets with the TF-targeted antisense RNA can limit the effects of IBMIR, thereby prolong NICC graft survival in SCID model.