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Oxygen Perfusion (Persufflation) of Human Pancreata Enhances Insulin Secretion and Attenuates Islet Proinflammatory Signaling

Kelly, Amy C., PhD1; Smith, Kate E., PhD2; Purvis, William G.3; Min, Catherine G., PhD2; Weber, Craig S.2; Cooksey, Amanda M.1; Hasilo, Craig4; Paraskevas, Steven, MD, PhD4; Suszynski, Thomas M., MD, PhD3; Weegman, Bradley P., PhD3; Anderson, Miranda J.1; Camacho, Leticia E., PhD1; Harland, Robert C., MD3; Loudovaris, Thomas, PhD3; Jandova, Jana, PhD3; Molano, Diana S.3; Price, Nicholas D.3; Georgiev, Ivan G.3; Scott, William E. III, PhD5; Manas, Derek M.D., PhD5; Shaw, James A.M., PhD5; O'Gorman, Doug6; Kin, Tatsuya, MD, PhD6; McCarthy, Fiona M., PhD1; Szot, Gregory L.7; Posselt, Andrew M., MD, PhD7; Stock, Peter G., MD, PhD7; Karatzas, Theodore, MD8; Shapiro, A.M. James, MD, PhD6; Lynch, Ronald M., PhD2; Limesand, Sean W., PhD1; Papas, Klearchos K., PhD3

doi: 10.1097/TP.0000000000002400
Original Basic Science—General

Background All human islets used in research and for the clinical treatment of diabetes are subject to ischemic damage during pancreas procurement, preservation, and islet isolation. A major factor influencing islet function is exposure of pancreata to cold ischemia during unavoidable windows of preservation by static cold storage (SCS). Improved preservation methods may prevent this functional deterioration. In the present study, we investigated whether pancreas preservation by gaseous oxygen perfusion (persufflation) better preserved islet function versus SCS.

Methods Human pancreata were preserved by SCS or by persufflation in combination with SCS. Islets were subsequently isolated, and preparations in each group matched for SCS or total preservation time were compared using dynamic glucose-stimulated insulin secretion as a measure of β-cell function and RNA sequencing to elucidate transcriptomic changes.

Results Persufflated pancreata had reduced SCS time, which resulted in islets with higher glucose-stimulated insulin secretion compared to islets from SCS only pancreata. RNA sequencing of islets from persufflated pancreata identified reduced inflammatory and greater metabolic gene expression, consistent with expectations of reducing cold ischemic exposure. Portions of these transcriptional responses were not associated with time spent in SCS and were attributable to pancreatic reoxygenation. Furthermore, persufflation extended the total preservation time by 50% without any detectable decline in islet function or viability.

Conclusions These data demonstrate that pancreas preservation by persufflation rather than SCS before islet isolation reduces inflammatory responses and promotes metabolic pathways in human islets, which results in improved β cell function.

1 School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, AZ.

2 Physiological Sciences, University of Arizona, Tucson, AZ.

3 Department of Surgery, Institute for Cellular Transplantation, University of Arizona, Tucson, AZ.

4 Human Islet Transplant Laboratory, McGill University Health Centre, Montreal, Quebec, Canada.

5 Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.

6 Clinical Islet Transplant Program, Alberta Diabetes Institute, University of Alberta, Edmonton, Alberta, Canada.

7 Department of Surgery, University of California San Francisco, San Francisco, CA.

8 Medical School, University of Athens, Athens, Greece.

Received 1 March 2018. Revision received 4 June 2018.

Accepted 19 June 2018.

This work was supported by the National Institutes of Health grants NIH/SBIR 5R44DK070400-04 (K.K.P., Co-Principal Investigator with Dr. Linda Templeman, Giner Inc.) and R01DK084842 (S.W.L., Principal Investigator) and NIH/NIDDK DP3 grant 1DP3DK106933-01 (K.K.P., Principal Investigator).

Duality of Interest: Klearchos K. Papas, PhD served as Consultant/ Chair SAB Giner Inc, is a co-inventor on pending patents related to persufflation with University of Minnesota and Giner Inc., and was the PI or co-PI on the NIH/NIDDK grants that partially funded the work. He is no longer affiliated with Giner Inc. "The remaining authors declare no conflicts of interest".

Guarantee: K.K.P. is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

K.K.P., A.C.K., K.E.S., S.W.L, and R.M.L. designed the study. Pancreas persufflation and islet isolations were conducted by C.G.M., J.J., K.E.S., N.D.P, D.S.M., I.G.G., T.L., C.P.H, S.P., W.E.S., D.M., J.A.S., D.O., T.K., G.L.S., A.M.P., P.G.S., T.K., A.M.J.S., B.P.W., and R.C.H. Functional islet assessments at the University of Arizona were conducted by K.E.S., C.W., W.G.P., C.G.M., D.S.M., and N.D.P. Technical aspects of sequencing and subsequent data analysis were conducted by A.C.K., A.M.C, L.E.C., F.M.M., R.M.L., and S.W.L. Traditional RNA quantification was conducted by A.C.K., K.E.S., L.E.C., M.J.A. All authors contributed to data interpretation, reviewed the article and approved the final version.

Correspondence: Klearchos K. Papas, PhD, Department of Surgery, Institute for Cellular Transplantation, University of Arizona, Medical Research Bldg, 1656 E. Mabel St, Rm 122, Tucson, AZ 85724. (kkpapas@surgery.arizona.edu).

Supplemental digital content (SDC) is available for this article. Direct URL citations appear in the printed text, and links to the digital files are provided in the HTML text of this article on the journal’s Web site (www.transplantjournal.com).

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