PARP-1 Inhibition Rescues Short Lifespan in Hyperglycemic C. Elegans And Improves GLP-1 Secretion in Human Cells
Xia Qianghua1, Lu Sumei1, Ostrovsky Julian1, McCormack Shana E2,3,4, Falk Marni J1,3, Grant Struan F. A1,2,3,4,*
1Division of Human Genetics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA. 2Division of Endocrinology and Diabetes, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA. 3Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. 4Institute of Diabetes, Obesity and Metabolism, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
TCF7L2 is located at one of the most strongly associated type 2 diabetes loci reported to date. We previously reported that the most abundant member of a specific protein complex to bind across the presumed causal variant at this locus, rs7903146, was poly [ADP-ribose] polymerase type 1 (PARP-1). We analyzed the impact of PARP-1 inhibition on C. elegans health in the setting of hyperglycemia and on glucose-stimulated GLP-1 secretion in human intestinal cells. Given that high glucose concentrations progressively shorten the lifespan of C. elegans, in part by impacting key well-conserved insulin-modulated signaling pathways, we investigated the effect of PARP-1 inhibition with Olaparib on the lifespan of C. elegans nematodes under varying hyperglycemic conditions. Subsequently, we investigated whether Olaparib treatment had any effect on glucose-stimulated GLP-1 secretion in the human NCI-H716 intestinal cell line, a model system for the investigation of enteroendocrine function. Treatment with 100uM Olaparib in nematodes exposed to high concentrations of glucose led to significant lifespan rescue. The beneficial lifespan effect of Olaparib appeared to require both PARP-1 and TCF7L2, since treatment had no effect in hyperglycemic conditions in knock-out worm strains for either of these homologs. Further investigation using the NCI-H716 cells revealed that Olaparib significantly enhanced secretion of the incretin, GLP-1, plus the gene expression of TCF7L2, GCG and PC1. These data from studies in both C. elegans and a human cell line suggest that PARP-1 inhibition offers a novel therapeutic avenue to treat type 2 diabetes.
Xia Qianghua,Lu Sumei,Ostrovsky Julian, et al. PARP-1 Inhibition Rescues Short Lifespan in Hyperglycemic C. Elegans And Improves GLP-1 Secretion in Human Cells[J]. Aging and disease,
2018, 9(1): 17-30.
Figure 1. High glucose conditions shorten C. elegans lifespan
Wild-type N2 C. elegans cultured under standard and high glucose conditions and subsequent lifespan assays. (A) Determination of lifespan for N2 worms unexposed or exposed to 16 mM glucose (P<0.002, log-rank test, n=100); (B) Determination of lifespan for N2 worms unexposed or exposed to 25 mM glucose (P<0.0001, log-rank test, n=100).
Figure 2. The lifespan-shortening effects of high glucose reduced by Olaparib treatment
(A) Determination of lifespan for N2 worms exposed to 16 mM glucose together with 100 uM or 0 µM Olaparib (P<0.0001, log-rank test, n=100); (B) Determination of lifespan for N2 worms exposed to 25 mM glucose together with 100 uM or 0 µM Olaparib (P<0.05, log-rank test, n=100).
Figure 3. The beneficial effect of Olaparib treatment on C. elegans lifespan in the setting of high glucose was target-specific and dependent on the PARP-1 related signaling pathway
(A) Determination of lifespan for PARP-1 homolog, pme-1, worms exposed to 16 mM glucose together with 100 µM or without Olaparib (P>0.05, log-rank test, n=100); (B) Determination of lifespan for pme-1 worms exposed to 25 mM glucose together with 100 µM or 0 µM Olaparib (P>0.05, log-rank test, n=100).
Figure 4. TCF7L2 homolog, pop-1, is required for the beneficial effect in the setting of high glucose of Olaparib treatment on lifespan
(A) Determination of lifespan for N2 worms fed with HT115 feeding strain containing the pL4440 empty vector and exposed to 16 mM glucose with 100 µM or 0 uM Olaparib (P<0.03, log-rank test, n=100); (B) Determination of lifespan for N2 worms fed with HT115 feeding strain containing the pL4440 pL4440/pop-1 RNAi and exposed to 16 mM glucose with 100uM or 0 µM Olaparib (P>0.05, log-rank test, n=100).
Figure 5. Olaparib enhances promotes GLP-1 secretion in NCI-H716 cells
Cells were stimulated for 30 minutes with or without 16 mM glucose. GLP-1 was measured by ELISA. Bars represent the mean of three independent experiments normalized to the control. Error bars indicate standard deviation. Statistical analyses were performed by two-tailed Student’s t-test and significance is denoted by asterisks where *P<0.05.
Figure 6. Olaparib treatment of endoendocrine intestinal cells increases expression of (A)TCF7L2, (B)GCG and (C)PC1
Relative levels of individual gene expression in the control and the Olaparib treatment groups determined by quantitative PCR with GAPDH normalization. Values are the mean of three independent experiments. Error bars indicate standard deviation. Statistical analyses were performed by two-tailed Student’s t-test and significance is denoted by asterisks where *P<0.05 or **P<0.01.
Figure 7. Integrated model to define the role of PARP-1 inhibition in the type 2 diabetes related pathway
Olaparib treatment increases the expression of TCF7L2, GCG and PC1, thereby enhancing GLP-1 secretion in the endoendocrine L type gastrointestinal cells and subsequently promoting insulin secretion in the pancreatic β-cell.
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