Pancreatic cell tracing, lineage tagging and targeted genetic manipulations in multiple cell types using pancreatic ductal infusion of adeno-associated viral vectors and/or cell-tagging dyes

Xiao, Xianmin; Guo, Ping; Prasadan, Krishna; Shiota, Chiyo; Peirish, Lauren; Fischbach, Shane; Song, Zewen; Gaffar, Iljana; Wiersch, John; El-Gohary, Yousef; Husain, Sohail Z.; Gittes, George K.

doi:10.1038/nprot.2014.183

Cited by 66 publications

(92 citation statements)

References 24 publications

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“…Only male mice were used in the current experiments. Pancreatic intraductal virus infusion was performed as described before (36). AAV8 viruses (titration of 10 12 genome copy particles/ml and in a volume of 150 l) were delivered via catheter at a rate of 6 l/min.…”

Section: Methodsmentioning

confidence: 99%

Forkhead Box Protein 1 (FoxO1) Inhibits Accelerated β Cell Aging in Pancreas-specific SMAD7 Mutant Mice

Xiao¹,

Chen

Guo

et al. 2017

Journal of Biological Chemistry

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Edited by Jeffrey E. PessinThe mechanisms underlying the effects of exocrine dysfunction on the development of diabetes remain largely unknown. Here we show that pancreatic depletion of SMAD7 resulted in age-dependent increases in ␤ cell dysfunction with accelerated glucose intolerance, followed by overt diabetes. The accelerated ␤ cell dysfunction and loss of proliferation capacity, two features of ␤ cell aging, appeared to be non-cell-autonomous, secondary to the adjacent exocrine failure as a "bystander effect." Increased Forkhead box protein 1 (FoxO1) acetylation and nuclear retention was followed by progressive FoxO1 loss in ␤ cells that marked the onset of diabetes. Moreover, forced FoxO1 expression in ␤ cells prevented ␤ cell dysfunction and loss in this model. Thus, we present a model of accelerated ␤ cell aging that may be useful for studying the mechanisms underlying ␤ cell failure in diabetes. Moreover, we provide evidence highlighting a critical role of FoxO1 in maintaining ␤ cell identity in the context of SMAD7 failure.␤ Cell dysfunction and failure are hallmarks of type 2 diabetes (1-4). Oxidative stress, endoplasmic reticulum stress, hypoxic stress, and cytokine toxicity can all lead to ␤ cell apoptosis, autophagy, and replication defects, resulting in global ␤ cell failure (3, 4). Thus, ␤ cell replacement may be an ideal strategy for treating diabetes (5). Although ␤ cell neogenesis from non-␤ cell sources has been extensively studied, most evidence suggests that postnatal ␤ cell expansion predominantly results from ␤ cell replication (6 -11). However, normally, the ␤ cell replication ratio is fairly low in young adult mice and then gets progressively lower with increasing age (4, 12-16). Meanwhile, recent studies strongly suggest that protection of the differentiated phenotype of existing ␤ cells is critical for the maintenance of a functional ␤ cell mass and for the prevention of type 2 diabetes (2,(17)(18)(19), in which the transcription factor Forkhead box protein 1 (FoxO1) 3 appears to play a key protective role against ␤ cell senescence and failure.TGF␤ superfamily signaling pathways are essential for pancreas development (20), postnatal ␤ cell homeostasis, and proper function (21-23). Binding of TGF␤ superfamily ligands to a type II receptor catalyzes the phosphorylation of a type I receptor to trigger downstream signaling cascades. SMAD7 is a general inhibitor of all TGF␤ superfamily pathways, and we have shown recently that SMAD7 plays a critical role during pancreas development and ␤ cell replication (20,(22)(23)(24). Here we studied ␤ cell function in a model of pancreatic SMAD7 depletion (SMAD7 Ptf1a ). ResultsPancreas-specific (SMAD7 Ptf1a ) SMAD7 Knockout Mice-To characterize the role of SMAD7 in ␤ cell development and postnatal ␤ cell mass homeostasis, we deleted pancreatic SMAD7 by crossing SMAD7fx/fx mice with Ptf1a-Cre mice to generate Ptf1a-Cre; SMAD7fx/fx mice, simplified as SMAD7 Ptf1a (supplemental Fig. 1, A and B). In Ptf1a-Cre mice, in which SMAD7 was deleted in the major...

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Section: Methodsmentioning

confidence: 99%

Forkhead Box Protein 1 (FoxO1) Inhibits Accelerated β Cell Aging in Pancreas-specific SMAD7 Mutant Mice

Xiao¹,

Chen

Guo

et al. 2017

Journal of Biological Chemistry

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“…The MIP-GFP mouse pancreas was first perfused with 0.125 mg/ml LiberaseTL (Roche, Nutley, NJ, USA) from the common bile duct, as has been described in published protocol [23,24], then was incubated in 0.125 mg/ml LiberaseTL in a 37°C shaker at 200rpm for 45 minutes. Histopaque1077 and Histopaque1119 (Sigma-Aldrich) were mixed at a ratio of 9:33 (vol:vol) to generate Histopaque1110.…”

Section: Methodsmentioning

confidence: 99%

Pre-Eclampsia-Associated Reduction in Placental Growth Factor Impaired Beta Cell Proliferation Through PI3k Signalling

Ying

Cai

et al. 2015

Cell Physiol Biochem

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Background/Aim: Reduction in serum placental growth factor (PLGF) frequently co-occurs with preeclampsia (PE) and gestational diabetes mellitus (GDM). Recently, we reported that impairment in gestational beta-cell mass growth may result from PE-associated reduction in PLGF and lead to development of GDM. Here, we studied the underlying mechanisms. Methods: We co-cultured primary mouse beta cells with mouse islet endothelial cells (MS1), with or without PLGF. We also cultured beta cells in conditioned media from PLGF-treated MS1. Specific signal-pathway inhibitors were applied to cultured beta cells in conditioned media from PLGF-treated MS1. We analysed beta-cell proliferation by BrdU incorporation. We analysed changes in cell number by a MTT assay. We analysed protein levels of cell-cycle regulators in beta cells by Western blot. Results: PLGF itself failed to induce beta-cell proliferation, but significantly augmented proliferation of beta cells co-cultured with MS1, which resulted in significant increases in cell number. Conditioned media from the PLGF-treated MS1 cells similarly induced beta-cell proliferation, which was abolished by inhibition of PI3k/Akt signalling, but not by inhibition of either ERK/MAPK or JNK signalling. The induction of beta-cell proliferation by PLGF-treated MS1 cells appeared to involve decreases in cell-cycle inhibitors p21 and p27, and increases in cell-cycle activators CDK4 and CyclinD1. Conclusion: Gestational PLGF may target islet endothelial cells to release growth factors that activate PI3k/Akt signalling in beta cells to increase their proliferation. PE-associated reduction in PLGF impairs these processes to result in GDM.

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“…AAV6 and AAV8 appear to have the highest infection efficiency in the pancreas compared with other AAV serotypes and transduce pancreatic acinar cells, islets, and ducts (27,46). In previous work, intrapancreatic ductal infusion of AAVs successfully yielded pancreas-specific infectivity (27,46,47).…”

Section: Discussionmentioning

confidence: 92%

Dynamic Imaging of Pancreatic Nuclear Factor κB (NF-κB) Activation in Live Mice Using Adeno-associated Virus (AAV) Infusion and Bioluminescence

Orabi

Sah

Javed

et al. 2015

Journal of Biological Chemistry

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Background: NF-B is an important signaling molecule in the development of acute pancreatitis. Results: Adeno-associated virus-NF-B-luciferase-infused mice showed a 77-and 140-fold increase in pancreas-specific NF-B bioluminescence following caerulein and caerulein ϩ LPS pancreatitis, respectively. Conclusion: NF-B activation can be examined in a live, dynamic fashion during pancreatic inflammation. Significance: This technique offers a valuable tool to study real-time activation of NF-B in vivo.

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Pancreatic cell tracing, lineage tagging and targeted genetic manipulations in multiple cell types using pancreatic ductal infusion of adeno-associated viral vectors and/or cell-tagging dyes

Cited by 66 publications

References 24 publications

Forkhead Box Protein 1 (FoxO1) Inhibits Accelerated β Cell Aging in Pancreas-specific SMAD7 Mutant Mice

Forkhead Box Protein 1 (FoxO1) Inhibits Accelerated β Cell Aging in Pancreas-specific SMAD7 Mutant Mice

Pre-Eclampsia-Associated Reduction in Placental Growth Factor Impaired Beta Cell Proliferation Through PI3k Signalling

Dynamic Imaging of Pancreatic Nuclear Factor κB (NF-κB) Activation in Live Mice Using Adeno-associated Virus (AAV) Infusion and Bioluminescence

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