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Insulin Action/Molecular Metabolism INSULIN ACTION—ADIPOCYTECATEGORY BIOLOGY 1692-P 1694-P Stress in Beta Cells Obtained with Laser Capture Microdissection Biopsy-Proven Insulitis of Clinical Islet Transplantation Is Not from Cadaver Pancreases of Brain Dead Donors Reversed by Steroid Therapy AREF EBRAHIMI, MIN-HO JUNG, JONATHAN M. DREYFUSS, HUI PAN, DENNIS ANNA LAM, BEHRUZ RAHIMI, SHARLEEN IMES, KIM SOLEZ, JAMES SHAPIRO, C. SGROI, SUSAN BONNER-WEIR, GORDON C. WEIR, Boston, MA PETER A. SENIOR, Edmonton, AB, Canada Brain death of pancreas donors is thought to lead to the expression of Gradual decline in islet function remains a challenge in clinical islet infl ammatory, stress and apoptotic pathways in isolated islets resulting in transplantation (CIT), but acute graft loss is relatively uncommon. Here we poor clinical outcomes. To test this hypothesis we obtained cadaveric pan- describe a case of acute decline in graft function with histology suggesting creases from brain dead pancreatic donors (n=7, mean age 5011) and normal an immune mechanism. A 49 year old female (BMI 24.4 kg/m2, insulin 0.3 pancreatic tissue obtained at surgery done for pancreatic neoplasms (n=7, U/kg) with type 1 diabetes for 37 years underwent two CIT (6071 and 6827 age 699). Frozen sections were subjected to laser capture microdissection islet equivalents/kg) following alemtuzumab induction with tacrolimus (TAC, to obtain beta-cell rich islet tissue, from which extracted RNA was analyzed mean 8 ug/L) and mycophenolate mofetil for maintenance. Initial engraft- with Affymetrix arrays. Gene expression of the two groups was evaluated ment was reasonable (β2 score of 12 at 1 week), but β2 score gradually with principle component analysis (PCA), and differential expression anal- declined (Figure 1) rising to 19 after the second CIT. Insulin independence ysis was performed for genes and pathways. The pathways expressed at was achieved only briefl y following each CIT. There was an acute decline the highest signifi cance included Glycolysis, Unfolded Protein Response, in β2 score after day 120 (Figure 1) prompting liver biopsy. Three collections MTORC1 Signaling, and Pancreatic Beta Cells. Pathways of Apoptosis were of insulin staining cells heavily infi ltrated with mononuclear infl ammatory not differentially expressed. cells and eosinophils were seen. TAC was increased (mean 14 ug/L) and A striking fi nding was the unfolded protein response (UPR), which is a pro- prednisone 50 mg/day started, but graft function did not recover. Pre-CIT tective component of the endoplasmic reticulum (ER) stress response that anti-insulin antibodies were positive and anti-GAD negative, while panel if severe can cause apoptosis. An important group of protective chaperone reactive antibodies were 0% pre- and post-CIT. This is the fi rst report of genes were upregulated in the cadaver donors (HSP90B1, HSPA5, PDIA6, biopsy proven insulitis post-CIT, which may be due to acute rejection and/ DnaJB9 and DnaJC3). In addition the protective genes of ER-associated pro- or recurrent autoimmunity. Unfortunately, this did not reverse with steroid tein degradation (ERAD) proteins (DERL 1, 2, 3) were upregulated. In con- therapy. Further study of allo- and auto-immunity in acute CIT failure may trast the proapoptotic “executioner” genes CHOP and JNK were not upregu- allow targeted immune therapy. lated. Other evidence of stress included upregulation of genes associated Figure 1. Post-CIT Graft Function as Measured by BETA-2 Score, a Clinical with NFkB activation and TNF pathways, including TNFRSF1A, IL17RB and Composite Score (Fasting C-peptide, Fasting Blood Glucose, Insulin Dose, IL13RA1. Other upregulated genes of interest included REG1B, LDHA and Tis- and HbA1C) after Two Transplants. sue Factor. In conclusion, while there was little evidence of active apoptosis in beta cells from cadaver donors, stress markers were found that could represent vulnerability resulting from brain death and/or the trauma of organ preser- vation. This could account for some of the early beta cell death found with islet transplantation. Supported By: National Institutes of Health 1693-P Abnormal Responses to Oral and Intravenous Glucose Stimula- tion in Intestinal Transplant Recipients With or Without Pancreas Allograft DAHAE LEE, LAURENS CEULEMANS, DANIEL JACOBS-TULLENEERS-THEVISSEN, BART KEYMEULEN, ILSE WEETS, DIETHARD MONBALIU, JACQUES PIRENNE, CHANTAL MATHIEU, PIETER GILLARD, Leuven, Belgium, Jette, Belgium Technique of liver-small intestinal transplantation (LITx) often includes a partial or whole pancreas allograft to preserve bile duct continuity. Aim of the study was to measure responses to oral and intravenous (IV) glucose stimulation in LITx recipients in comparison to recipients of isolated small intestinal transplant (ITx) and nondiabetic controls. Plasma glucose, C-peptide and insulin were measured during 180-minute oral glucose tolerance test (OGTT) and 160-minute hyperglycemic clamp (HG clamp) in nondiabetic LITx (n = 3) and ITx (n = 3) recipients under immuno- INSULIN ACTION—ADIPOCYTE BIOLOGY suppression with azathioprine, tacrolimus and low-dose corticosteroids. Ten POSTERS healthy volunteers served as controls. Insulin Action/ All but 1 LITx and ITx recipients showed abnormal glucose profi le after Moderated Poster Discussion: Feel the Burn—Factors that Activate OGTT despite higher basal C-peptide. 2/3 ITx recipients showed higher glu- Molecular Metabolism Brown Fat (Posters: 1695-P to 1701-P), see page 14. cose peak during OGTT and 45% lower functional beta cell mass (FBM) than controls. In 1 LITx recipient with partial pancreas allograft, OGTT resulted in & 1695-P delayed but higher glucose peak in parallel with high C-peptide and insulin Differential Roles of Fox0 Proteins in White and Brown Adipose levels, leading to rapid decrease in glucose and severe hypoglycemia. Both Tissue LITx recipients with partial pancreas allograft had FBM above percentile 85 ERICA HOMAN, BRIAN T. O’NEILL, MASAJI SAKAGUCHI, CHRISTIE PENNIMAN, of controls. One LITx recipient with whole pancreas allograft showed very DOMENICO ACCILI, C. RONALD KAHN, Boston, MA, New York, NY high baseline C-peptide, due to systemic venous drainage of recipient pan- Insulin and IGF-1 are essential for mediating normal adipocyte differen- creas and high insulin resistance (HOMA-R=3.60). Glucose peak during OGTT tiation and metabolism. In insulin/IGF responsive tissues, insulin and IGF-1 was higher and delayed but without late hypoglycemia. During all phases induce phosphorylation of the Forkhead Box O family of transcription factors of the HG clamp, C-peptide and insulin were higher with FBM calculated at (Fox0s) via action of Akt maintaining their cytoplasmic localization, reversing 150% of controls. the activation of multiple Fox0-responsive genes involved in maintenance of Intestinal transplant recipients show abnormal responses to both oral energy homeostasis, glucose metabolism, cell cycle arrest, and cell death. and IV glucose stimulation. Several factors may explain these observations: We explored whether Fox0 activation controls the decrease in white and intestinal denervation, loss of normal anatomy, loss of FBM and insulin resis- brown fat mass, glucose intolerance, and decrease in brown fat activation tance. Analysis of these complex cases can help to understand regulation of observed in mice with fat-specifi c knockout of both the insulin and IGF-1 glucose metabolism in normal and posttransplant conditions. receptors (FIGIRKO mice). To this end, we generated mice with fat-specifi c knockout of the insulin and IGF-1 receptors, as well as knock-out of the three Fox0s expressed in fat [Fox01, Fox03, and Fox04] (F-Quint KO) and compared these with control and FIGIRKO mice. As previously observed, FIGIRKO mice For author disclosure information, see page A696. & Moderated Poster Discussion ADA-Supported Research A440 INSULIN ACTION—ADIPOCYTECATEGORY BIOLOGY exhibited a marked loss of all white and brown fat depots, marked hypergly- & 1698-P cemia, insulin resistance, fatty liver and inability to maintain body tempera- Raptor Defi ciency Promotes Browning of White Adipose Tissue via ture during a cold challenge. By contrast, the F-Quint KO mice had similar glu- Adiponectin-dependent Mechanisms cose tolerance to control animals, had normalized brown fat mass and were XIAOFENG DING, YAN LUO, XIN YANG, XIAOQING WANG, XIANYUN ZHENG, able to maintain body temperature during cold exposure. However, like the XUEXIAN YANG, MEILIAN LIU, Albuquerque, NM, Changsha, China FIGIRKO mice, the F-Quint KO mice failed to respond to insulin stimulation, Beige adipocytes burn lipid by dissipating energy in the form of heat and had markedly decreased subcutaneous fat pad weights, absence of perigo- offer a new way to battle obesity and its related disorders. However, what nadal fat pads, and hepatosteatosis. Thus, we demonstrate differential intracellular signaling pathways drive the browning effect remains largely regulation of Fox0 proteins in adipose tissue such that suppression of Fox0s unknown. Here, we show that inhibition of mammalian target of rapamycin by insulin/IGF-1 action is critical for brown fat differentiation and systemic complex 1 (mTORC1) by adipose specifi c-deletion of raptor (raptor fKO), a glucose metabolism, but not critical for the regulation of white adipose tis- key component of mTORC1 promoted browning of inguinal WAT (iWAT) and sue mass and systemic insulin sensitivity. enhanced basal and cold-induced energy expenditure, concurrently with an Supported By:
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