Islet organoid as a promising model for diabetes

Abstract

Studies on diabetes have long been hampered by a lack of authentic disease models that, ideally, should be unlimited and able to recapitulate the abnormalities involved in the development, structure, and function of human pancreatic islets under pathological conditions. Stem cell-based islet organoids faithfully recapitulate islet development invitroand provide large amounts of three-dimensional functional islet biomimetic materials with a morphological structure and cellular composition similar to those of native islets. Thus, islet organoids hold great promise for modeling islet development and function, deciphering the mechanisms underlying the onset of diabetes, providing an invitrohuman organ model for infection of viruses such as SARS-CoV-2, and contributing to drug screening and autologous islet transplantation. However, the currently established islet organoids are generally immature compared with native islets, and further efforts should be made to improve the heterogeneity and functionality of islet organoids, making it an authentic and informative disease model for diabetes. Here, we review the advances and challenges in the generation of islet organoids, focusing on human pluripotent stem cell-derived islet organoids, and the potential applications of islet organoids as disease models and regenerative therapies for diabetes.

Keywords

islet organoid / diabetes / pluripotent/adult stem cell / pancreatic β cell / disease model

References

[1] Aamodt KI, Powers AC (2017) Signals in the pancreatic islet microenvironment influence beta-cell proliferation. Diabetes Obes Metab 19(Suppl 1):124–136

[2] Aguayo-Mazzucato C, Bonner-Weir S (2018) Pancreatic beta cell regeneration as a possible therapy for diabetes. Cell Metab 27:57–67

[3] Agulnick AD, Ambruzs DM, Moorman MA, Bhoumik A, Cesario RM, Payne JK, Kelly JR, Haakmeester C, Srijemac R, Wilson AZ (2015) Insulin-producing endocrine cells differentiated in vitro from human embryonic stem cells function in macroencapsulation devices in vivo. Stem Cells Transl Med 4:1214–1222

[4] Ahren B (2000) Autonomic regulation of islet hormone secretionimplications for health and disease. Diabetologia 43:393–410

[5] Alvarez-Dominguez JR, Donaghey J, Rasouli N, Kenty JHR, Helman A, Charlton J, Straubhaar JR, Meissner A, Melton DA (2020) Circadian entrainment triggers maturation of human in vitro islets. Cell Stem Cell 26(108–122):e110

[6] Ameri J, Borup R, Prawiro C, Ramond C, Schachter KA, Scharfmann R, Semb H (2017) Efficient generation of glucose-responsive beta cells from isolated GP2(+) human pancreatic progenitors. Cell Rep 19:36–49

[7] Amin S, Cook B, Zhou T, Ghazizadeh Z, Lis R, Zhang T, Khalaj M, Crespo M, Perera M, Xiang JZ (2018) Discovery of a drug candidate for GLIS3-associated diabetes. Nat Commun 9:2681

[8] Arrojo e Drigo R, Ali Y, Diez J, Srinivasan DK, Berggren PO, Boehm BO (2015) New insights into the architecture of the islet of Langerhans: a focused cross-species assessment. Diabetologia 58:2218–2228

[9] Asahara S, Etoh H, Inoue H, Teruyama K, Shibutani Y, Ihara Y, Kawada Y, Bartolome A, Hashimoto N, Matsuda T (2015) Paternal allelic mutation at the Kcnq1 locus reduces pancreatic beta-cell mass by epigenetic modification of Cdkn1c. Proc Natl Acad Sci USA 112:8332–8337

[10] Ashcroft FM, Rorsman P (2012) Diabetes mellitus and the beta cell: the last ten years. Cell 148:1160–1171

[11] Augsornworawat P, Maxwell KG, Velazco-Cruz L, Millman JR (2020) Single-cell transcriptome profiling reveals beta cell maturation in stem cell-derived islets after transplantation. Cell Rep 32:108067

[12] Bader E, Migliorini A, Gegg M, Moruzzi N, Gerdes J, Roscioni SS, Bakhti M, Brandl E, Irmler M, Beckers J (2016) Identification of proliferative and mature beta-cells in the islets of Langerhans. Nature 535:430–434

[13] Bakhti M, Scheibner K, Tritschler S, Bastidas-Ponce A, Tarquis-Medina M, Theis FJ, Lickert H (2019) Establishment of a highresolution 3D modeling system for studying pancreatic epithelial cell biology in vitro. Mol Metab 30:16–29

[14] Balboa D, Saarimaki-Vire J, Borshagovski D, Survila M, Lindholm P, Galli E, Eurola S, Ustinov J, Grym H, Huopio H (2018) Insulin mutations impair beta-cell development in a patient-derived iPSC model of neonatal diabetes. Elife7

[15] Basford CL, Prentice KJ, Hardy AB, Sarangi F, Micallef SJ, Li X, Guo Q, Elefanty AG, Stanley EG, Keller G (2012) The functional and molecular characterisation of human embryonic stem cellderived insulin-positive cells compared with adult pancreatic beta cells. Diabetologia 55:358–371

[16] Bi H, Ye K, Jin S (2020) Proteomic analysis of decellularized pancreatic matrix identifies collagen V as a critical regulator for islet organogenesis from human pluripotent stem cells. Biomaterials 233:119673

[17] Bishay RH, Greenfield JR (2016) A review of maturity onset diabetes of the young (MODY) and challenges in the management of glucokinase-MODY. Med J Aust 205:480–485

[18] Borden P, Houtz J, Leach SD, Kuruvilla R (2013) Sympathetic innervation during development is necessary for pancreatic islet architecture and functional maturation. Cell Rep 4:287–301

[19] Bornstein SR, Rubino F, Khunti K, Mingrone G, Hopkins D, Birkenfeld AL, Boehm B, Amiel S, Holt RIG, Skyler JS (2020) Practical recommendations for the management of diabetes in patients with COVID-19. Lancet Diabetes Endocrinol 8:546–550

[20] Brissova M, Fowler MJ, Nicholson WE, Chu A, Hirshberg B, Harlan DM, Powers AC (2005) Assessment of human pancreatic islet architecture and composition by laser scanning confocal microscopy. J Histochem Cytochem 53:1087–1097

[21] Broda TR, McCracken KW, Wells JM (2019) Generation of human antral and fundic gastric organoids from pluripotent stem cells. Nat Protoc 14:28–50

[22] Bruin JE, Erener S, Vela J, Hu X, Johnson JD, Kurata HT, Lynn FC, Piret JM, Asadi A, Rezania A (2014) Characterization of polyhormonal insulin-producing cells derived in vitro from human embryonic stem cells. Stem Cell Res 12:194–208

[23] Cabrera O, Berman DM, Kenyon NS, Ricordi C, Berggrern PO, Caicedo A (2006) The unique cytoarchitecture of human pancreatic islets has implications for islet cell function. Proc Natl Acad Sci USA 103:2334–2339

[24] Cai EP, Ishikawa Y, Zhang W, Leite NC, Li J, Hou S, Kiaf B, Hollister-Lock J, Yilmaz NK, Schiffer CA (2020) Genome-scale in vivo CRISPR screen identifies RNLS as a target for beta cell protection in type 1 diabetes. Nat Metab 2:934–945

[25] Candiello J, Grandhi TSP, Goh SK, Vaidya V, Lemmon-Kishi M, Eliato KR, Ros R, Kumta PN, Rege K, Banerjee I (2018) 3D heterogeneous islet organoid generation from human embryonic stem cells using a novel engineered hydrogel platform. Biomaterials 177:27–39

[26] Cardenas-Diaz FL, Osorio-Quintero C, Diaz-Miranda MA, Kishore S, Leavens K, Jobaliya C, Stanescu D, Ortiz-Gonzalez X, Yoon C, Chen CS (2019) Modeling monogenic diabetes using human ESCs reveals developmental and metabolic deficiencies caused by mutations in HNF1A. Cell Stem Cell 25(273–289):e275

[27] Chen YJ, Finkbeiner SR, Weinblatt D, Emmett MJ, Tameire F, Yousefi M, Yang C, Maehr R, Zhou Q, Shemer R (2014) De novo formation of insulin-producing “neo-beta cell islets” from intestinal crypts. Cell Rep 6:1046–1058

[28] Choi KM, Seo YK, Yoon HH, Song KY, Kwon SY, Lee HS, Park JK (2008) Effect of ascorbic acid on bone marrow-derived mesenchymal stem cell proliferation and differentiation. J Biosci Bioeng 105:586–594

[29] Clevers H (2016) Modeling development and disease with organoids. Cell 165:1586–1597

[30] Coate KC, Cha J, Shrestha S, Wang W, Goncalves LM, Almaca J, Kapp ME, Fasolino M, Morgan A, Dai C (2020) SARS-CoV-2 cell entry factors ACE2 and TMPRSS2 are expressed in the microvasculature and ducts of human pancreas but are not enriched in beta cells. Cell Metab 32(1028–1040):e1024

[31] Cogger KF, Sinha A, Sarangi F, McGaugh EC, Saunders D, Dorrell C, Mejia-Guerrero S, Aghazadeh Y, Rourke JL, Screaton RA (2017) Glycoprotein 2 is a specific cell surface marker of human pancreatic progenitors. Nat Commun 8:331

[32] Cooper-Jones B, Ford C (2016) Islet cell replacement therapy for insulin-dependent diabetes. In: CADTH Issues in Emerging Health Technologies (Ottawa (ON)), pp 1–9

[33] D’Amour KA, Bang AG, Eliazer S, Kelly OG, Agulnick AD, Smart NG, Moorman MA, Kroon E, Carpenter MK, Baetge EE (2006) Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 24:1392–1401

[34] Davis JC, Alves TC, Helman A, Chen JC, Kenty JH, Cardone RL, Liu DR, Kibbey RG, Melton DA (2020) Glucose response by stem cell-derived beta cells in vitro is inhibited by a bottleneck in glycolysis. Cell Rep 31:107623

[35] de Vargas LM, Sobolewski J, Siegel R, Moss LG (1997) Individual beta cells within the intact islet differentially respond to glucose. J Biol Chem 272:26573–26577

[36] Dominguez-Bendala J, Qadir MMF, Pastori RL (2019) Pancreatic progenitors: there and back again. Trends Endocrinol Metab 30:4–11

[37] Dor Y, Brown J, Martinez OI, Melton DA (2004) Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation. Nature 429:41–46

[38] Dutta D, Heo I, Clevers H (2017) Disease modeling in stem cellderived 3D organoid systems. Trends Mol Med 23:393–410

[39] Dwivedi OP, Lehtovirta M, Hastoy B, Chandra V, Krentz NAJ, Kleiner S, Jain D, Richard AM, Abaitua F, Beer NL (2019) Loss of ZnT8 function protects against diabetes by enhanced insulin secretion. Nat Genet 51:1596–1606

[40] Eberhard D, Kragl M, Lammert E (2010) ‘Giving and taking’: endothelial and beta-cells in the islets of Langerhans. Trends Endocrinol Metab 21:457–463

[41] Fatehullah A, Tan SH, Barker N (2016) Organoids as an in vitro model of human development and disease. Nat Cell Biol 18:246–254

[42] Ghazizadeh Z, Kao DI, Amin S, Cook B, Rao S, Zhou T, Zhang T, Xiang Z, Kenyon R, Kaymakcalan O (2017) ROCKII inhibition promotes the maturation of human pancreatic beta-like cells. Nat Commun 8:298

[43] Giobbe GG, Crowley C, Luni C, Campinoti S, Khedr M, Kretzschmar K, De Santis MM, Zambaiti E, Michielin F, Meran L (2019) Extracellular matrix hydrogel derived from decellularized tissues enables endodermal organoid culture. Nat Commun 10:5658

[44] Gouzi M, Kim YH, Katsumoto K, Johansson K, Grapin-Botton A (2011) Neurogenin3 initiates stepwise delamination of differentiating endocrine cells during pancreas development. Dev Dyn 240:589–604

[45] Greggio C, De Franceschi F, Figueiredo-Larsen M, Gobaa S, Ranga A, Semb H, Lutolf M, Grapin-Botton A (2013) Artificial threedimensional niches deconstruct pancreas development in vitro. Development 140:4452–4462

[46] Guo D, Liu H, Ruzi A, Gao G, Nasir A, Liu Y, Yang F, Wu F, Xu G, Li YX (2017) Modeling congenital hyperinsulinism with ABCC8-deficient human embryonic stem cells generated by CRISPR/ Cas9. Sci Rep 7:3156

[47] Habener JF (2004) A perspective on pancreatic stem/progenitor cells. Pediatr Diabetes 5(Suppl 2):29–37

[48] Han X, Wang M, Duan S, Franco PJ, Kenty JH, Hedrick P, Xia Y, Allen A, Ferreira LMR, Strominger JL (2019) Generation of hypoimmunogenic human pluripotent stem cells. Proc Natl Acad Sci USA 116:10441–10446

[49] Han Y, Yang L, Duan X, Duan F, Nilsson-Payant BE, Yaron TM, Wang P, Tang X, Zhang T, Zhao Z (2020) Identification of candidate COVID-19 therapeutics using hPSC-derived lung organoids. bioRxiv

[50] Helman A, Cangelosi AL, Davis JC, Pham Q, Rothman A, Faust AL, Straubhaar JR, Sabatini DM, Melton DA (2020) A nutrientsensing transition at birth triggers glucose-responsive insulin secretion. Cell Metab 31(1004–1016):e1005

[51] Hogrebe NJ, Augsornworawat P, Maxwell KG, Velazco-Cruz L, Millman JR (2020) Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells. Nat Biotechnol 38:460–470

[52] Hrvatin S, O’Donnell CW, Deng F, Millman JR, Pagliuca FW, DiIorio P, Rezania A, Gifford DK, Melton DA (2014) Differentiated human stem cells resemble fetal, not adult, beta cells. Proc Natl Acad Sci USA 111:3038–3043

[53] Jennings RE, Berry AA, Strutt JP, Gerrard DT, Hanley NA (2015) Human pancreas development. Development 142:3126–3137

[54] Jiang K, Chaimov D, Patel SN, Liang JP, Wiggins SC, Samojlik MM, Rubiano A, Simmons CS, Stabler CL (2019) 3-D physiomimetic extracellular matrix hydrogels provide a supportive microenvironment for rodent and human islet culture. Biomaterials 198:37–48

[55] Johansson KA, Dursun U, Jordan N, Gu G, Beermann F, Gradwohl G, Grapin-Botton A (2007) Temporal control of neurogenin3 activity in pancreas progenitors reveals competence windows for the generation of different endocrine cell types. Dev Cell 12:457–465

[56] Kasputis T, Clough D, Noto F, Rychel K, Dye B, Shea LD (2018) Microporous polymer scaffolds for the transplantation of embryonic stem cell derived pancreatic progenitors to a clinically translatable site for the treatment of type I diabetes. ACS Biomater Sci Eng 4:1770–1778

[57] Kelly OG, Chan MY, Martinson LA, Kadoya K, Ostertag TM, Ross KG, Richardson M, Carpenter MK, D’Amour KA, Kroon E (2011) Cell-surface markers for the isolation of pancreatic cell types derived from human embryonic stem cells. Nat Biotechnol 29:750–756

[58] Kim Y, Kim H, Ko UH, Oh Y, Lim A, Sohn JW, Shin JH, Kim H, Han YM (2016) Islet-like organoids derived from human pluripotent stem cells efficiently function in the glucose responsiveness in vitro and in vivo. Sci Rep 6:35145

[59] Kroon E, Martinson LA, Kadoya K, Bang AG, Kelly OG, Eliazer S, Young H, Richardson M, Smart NG, Cunningham J (2008) Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol 26:443–452

[60] Kunisada Y, Tsubooka-Yamazoe N, Shoji M, Hosoya M (2012) Small molecules induce efficient differentiation into insulin-producing cells from human induced pluripotent stem cells. Stem Cell Res 8:274–284

[61] Kusmartseva I, Wu W, Syed F, Van Der Heide V, Jorgensen M, Joseph P, Tang X, Candelario-Jalil E, Yang C, Nick H (2020) Expression of SARS-CoV-2 entry factors in the pancreas of normal organ donors and individuals with COVID-19. Cell Metab 32(1041–1051):e1046

[62] Lamers MM, Beumer J, van der Vaart J, Knoops K, Puschhof J, Breugem TI, Ravelli RBG, Paul van Schayck J, Mykytyn AZ, Duimel HQ (2020) SARS-CoV-2 productively infects human gut enterocytes. Science 369:50–54

[63] Lancaster MA, Knoblich JA (2014) Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345:1247125

[64] Lee K, Cho H, Rickert RW, Li QV, Pulecio J, Leslie CS, Huangfu D (2019) FOXA2 is required for enhancer priming during pancreatic differentiation. Cell Rep 28(382–393):e387

[65] Leite NC, Sintov E, Meissner TB, Brehm MA, Greiner DL, Harlan DM, Melton DA (2020) Modeling type 1 diabetes in vitro using human pluripotent stem cells. Cell Rep 32:107894

[66] Lemper M, Leuckx G, Heremans Y, German MS, Heimberg H, Bouwens L, Baeyens L (2015) Reprogramming of human pancreatic exocrine cells to beta-like cells. Cell Death Differ 22:1117–1130

[67] Li M, Du W, Zhou M, Zheng L, Song E, Hou J (2018) Proteomic analysis of insulin secretory granules in INS-1 cells by protein correlation profiling. Biophys Rep 4:329–338

[68] Loomans CJM, Williams Giuliani N, Balak J, Ringnalda F, van Gurp L, Huch M, Boj SF, Sato T, Kester L, de Sousa Lopes SMC (2018) Expansion of adult human pancreatic tissue yields organoids harboring progenitor cells with endocrine differentiation potential. Stem Cell Rep 10:712–724

[69] Ma S, Viola R, Sui L, Cherubini V, Barbetti F, Egli D (2018) Beta cell replacement after gene editing of a neonatal diabetes-causing mutation at the insulin locus. Stem Cell Rep 11:1407–1415

[70] Mahaddalkar PU, Scheibner K, Pfluger S, Sterr M, Beckenbauer J, Irmler M, Beckers J, Knobel S, Lickert H (2020) Generation of pancreatic beta cells from CD177(+) anterior definitive endoderm. Nat Biotechnol 38:1061–1072

[71] Mamidi A, Prawiro C, Seymour PA, de Lichtenberg KH, Jackson A, Serup P, Semb H (2018) Mechanosignalling via integrins directs fate decisions of pancreatic progenitors. Nature 564:114–118

[72] Manzar GS, Kim EM, Zavazava N (2017) Demethylation of induced pluripotent stem cells from type 1 diabetic patients enhances differentiation into functional pancreatic beta cells. J Biol Chem 292:14066–14079

[73] Mattsson G (2005) The endothelial cells in islets of langerhans. Ups J Med Sci 110:1–15

[74] Maxwell KG, Augsornworawat P, Velazco-Cruz L, Kim MH, Asada R, Hogrebe NJ, Morikawa S, Urano F, Millman JR (2020) Geneedited human stem cell-derived beta cells from a patient with monogenic diabetes reverse preexisting diabetes in mice. Sci Transl Med12

[75] McGrath PS, Watson CL, Ingram C, Helmrath MA, Wells JM (2015) The basic helix-loop-helix transcription factor NEUROG3 is required for development of the human endocrine pancreas. Diabetes 64:2497–2505

[76] Meda P, Atwater I, Goncalves A, Bangham A, Orci L, Rojas E (1984) The topography of electrical synchrony among beta-cells in the mouse islet of Langerhans. Q J Exp Physiol 69:719–735

[77] Memon B, Karam M, Al-Khawaga S, Abdelalim EM (2018) Enhanced differentiation of human pluripotent stem cells into pancreatic progenitors co-expressing PDX1 and NKX6.1. Stem Cell Res Ther 9:15

[78] Millman JR, Xie C, Van Dervort A, Gurtler M, Pagliuca FW, Melton DA (2016) Generation of stem cell-derived beta-cells from patients with type 1 diabetes. Nat Commun 7:11463

[79] Miura K, Okada Y, Aoi T, Okada A, Takahashi K, Okita K, Nakagawa M, Koyanagi M, Tanabe K, Ohnuki M (2009) Variation in the safety of induced pluripotent stem cell lines. Nat Biotechnol 27:743–745

[80] Muniyappa R, Gubbi S (2020) COVID-19 pandemic, coronaviruses, and diabetes mellitus. Am J Physiol Endocrinol Metab 318:E736–E741

[81] Nair G, Hebrok M (2015) Islet formation in mice and men: lessons for the generation of functional insulin-producing beta-cells from human pluripotent stem cells. Curr Opin Genet Dev 32:171–180

[82] Nair GG, Liu JS, Russ HA, Tran S, Saxton MS, Chen R, Juang C, Li ML, Nguyen VQ, Giacometti S (2019) Recapitulating endocrine cell clustering in culture promotes maturation of human stem-cell-derived beta cells. Nat Cell Biol 21:263–274

[83] Nakagawa M, Takizawa N, Narita M, Ichisaka T, Yamanaka S (2010) Promotion of direct reprogramming by transformation-deficient Myc. Proc Natl Acad Sci USA 107:14152–14157

[84] Nessa A, Aziz QH, Thomas AM, Harmer SC, Tinker A, Hussain K (2015) Molecular mechanisms of congenital hyperinsulinism due to autosomal dominant mutations in ABCC8. Hum Mol Genet 24:5142–5153

[85] Nostro MC, Sarangi F, Yang C, Holland A, Elefanty AG, Stanley EG, Greiner DL, Keller G (2015) Efficient generation of NKX6-1+ pancreatic progenitors from multiple human pluripotent stem cell lines. Stem Cell Reports 4:591–604

[86] Nyeng P, Heilmann S, Lof-Ohlin ZM, Pettersson NF, Hermann FM, Reynolds AB, Semb H (2019) p120ctn-mediated organ patterning precedes and determines pancreatic progenitor fate. Dev Cell 49 (31–47):e39

[87] Oakie A, Li J, Fellows GF, Hess DA, Wang R (2018) Characterization and differentiation of sorted human fetal pancreatic ALDH(hi) and ALDH(hi)/CD133(+) cells toward insulin-expressing cells. Stem Cells Dev 27:275–286

[88] Ojaghi M, Soleimanifar F, Kazemi A, Ghollasi M, Soleimani M, Nasoohi N, Enderami SE (2019) Electrospun poly-l-lactic acid/ polyvinyl alcohol nanofibers improved insulin-producing cell differentiation potential of human adipose-derived mesenchymal stem cells. J Cell Biochem 120:9917–9926

[89] Okita K, Ichisaka T, Yamanaka S (2007) Generation of germlinecompetent induced pluripotent stem cells. Nature 448:313–317

[90] Oliver-Krasinski JM, Stoffers DA (2008) On the origin of the beta cell. Genes Dev 22:1998–2021

[91] Op de Beeck A, Eizirik DL (2016) Viral infections in type 1 diabetes mellitus–why the beta cells? Nat Rev Endocrinol 12:263–273

[92] O’Sullivan ES, Vegas A, Anderson DG, Weir GC (2011) Islets transplanted in immunoisolation devices: a review of the progress and the challenges that remain. Endocr Rev 32:827–844

[93] Pagliuca FW, Millman JR, Gürtler M, Segel M, Van Dervort A, Ryu JH, Peterson QP, Greiner D, Melton DA (2014) Generation of functional human pancreatic β cells in vitro. Cell 159:428–439

[94] Pathak S, Pham TT, Jeong JH, Byun Y (2019) Immunoisolation of pancreatic islets via thin-layer surface modification. J Control Release 305:176–193

[95] Pepper AR, Gala-Lopez B, Pawlick R, Merani S, Kin T, Shapiro AM (2015) A prevascularized subcutaneous device-less site for islet and cellular transplantation. Nat Biotechnol 33:518–523

[96] Petersen MBK, Goncalves CAC, Kim YH, Grapin-Botton A (2018) Recapitulating and deciphering human pancreas development from human pluripotent stem cells in a dish. Curr Top Dev Biol 129:143–190

[97] Phelps EA, Headen DM, Taylor WR, Thule PM, Garcia AJ (2013) Vasculogenic bio-synthetic hydrogel for enhancement of pancreatic islet engraftment and function in type 1 diabetes. Biomaterials 34:4602–4611

[98] Pictet RL, Clark WR, Williams RH, Rutter WJ (1972) An ultrastructural analysis of the developing embryonic pancreas. Dev Biol 29:436–467

[99] Qadir MMF, Alvarez-Cubela S, Klein D, Lanzoni G, Garcia-Santana C, Montalvo A, Placeres-Uray F, Mazza EMC, Ricordi C, Inverardi LA (2018) P2RY1/ALK3-expressing cells within the adult human exocrine pancreas are BMP-7 expandable and exhibit progenitor-like characteristics. Cell Rep 22:2408–2420

[100] Qadir MMF, Alvarez-Cubela S, Klein D, van Dijk J, Muniz-Anquela R, Moreno-Hernandez YB, Lanzoni G, Sadiq S, Navarro-Rubio B, Garcia MT (2020) Single-cell resolution analysis of the human pancreatic ductal progenitor cell niche. Proc Natl Acad Sci USA 117:10876–10887

[101] Ranjan AK, Joglekar MV, Hardikar AA (2009) Endothelial cells in pancreatic islet development and function. Islets 1:2–9

[102] Rezania A, Bruin JE, Riedel MJ, Mojibian M, Asadi A, Xu J, Gauvin R, Narayan K, Karanu F, O’Neil JJ (2012) Maturation of human embryonic stem cell-derived pancreatic progenitors into functional islets capable of treating pre-existing diabetes in mice. Diabetes 61:2016–2029

[103] Rezania A, Bruin JE, Xu J, Narayan K, Fox JK, O’Neil JJ, Kieffer TJ (2013) Enrichment of human embryonic stem cell-derived NKX6.1-expressing pancreatic progenitor cells accelerates the maturation of insulin-secreting cells in vivo. Stem Cells 31:2432–2442

[104] Rezania A, Bruin JE, Arora P, Rubin A, Batushansky I, Asadi A, O’Dwyer S, Quiskamp N, Mojibian M, Albrecht T (2014) Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat Biotechnol 32:1121–1133

[105] Rodriguez-Seguel E, Mah N, Naumann H, Pongrac IM, Cerda-Esteban N, Fontaine JF, Wang Y, Chen W, Andrade-Navarro MA, Spagnoli FM (2013) Mutually exclusive signaling signatures define the hepatic and pancreatic progenitor cell lineage diver-gence. Genes Dev 27:1932–1946

[106] Rohayem J, Ehlers C, Wiedemann B, Holl R, Oexle K, Kordonouri O, Salzano G, Meissner T, Burger W, Schober E (2011) Diabetes and neurodegeneration in Wolfram syndrome: a multicenter study of phenotype and genotype. Diabetes Care 34:1503–1510

[107] Romer AI, Singer RA, Sui L, Egli D, Sussel L (2019) Murine perinatal beta-cell proliferation and the differentiation of human stem cellderived insulin-expressing cells require NEUROD1. Diabetes 68:2259–2271

[108] Rosado-Olivieri EA, Anderson K, Kenty JH, Melton DA (2019) YAP inhibition enhances the differentiation of functional stem cellderived insulin-producing beta cells. Nat Commun 10:1464

[109] Rosado-Olivieri EA, Aigha II, Kenty JH, Melton DA (2020) Identification of a LIF-responsive, replication-competent subpopulation of human beta cells. Cell Metab 31(327–338):e326

[110] Roscioni SS, Migliorini A, Gegg M, Lickert H (2016) Impact of islet architecture on beta-cell heterogeneity, plasticity and function. Nat Rev Endocrinol 12:695–709

[111] Rossen NS, Anandakumaran PN, Zur Nieden R, Lo K, Luo W, Park C, Huyan C, Fu Q, Song Z, Singh-Moon RP (2020) Injectable therapeutic organoids using sacrificial hydrogels. iScience 23:101052

[112] Rukstalis JM, Habener JF (2007) Snail2, a mediator of epithelialmesenchymal transitions, expressed in progenitor cells of the developing endocrine pancreas. Gene Expr Patterns 7:471–479

[113] Russ HA, Parent AV, Ringler JJ, Hennings TG, Nair GG, Shveygert M, Guo T, Puri S, Haataja L, Cirulli V (2015) Controlled induction of human pancreatic progenitors produces functional beta-like cells in vitro. EMBO J 34:1759–1772

[114] Russell R, Carnese PP, Hennings TG, Walker EM, Russ HA, Liu JS, Giacometti S, Stein R, Hebrok M (2020) Loss of the transcription factor MAFB limits beta-cell derivation from human PSCs. Nat Commun 11:2742

[115] Saarimaki-Vire J, Balboa D, Russell MA, Saarikettu J, Kinnunen M, Keskitalo S, Malhi A, Valensisi C, Andrus C, Eurola S (2017) An activating STAT3 mutation causes neonatal diabetes through premature induction of pancreatic differentiation. Cell Rep 19:281–294

[116] Saeedi P, Petersohn I, Salpea P, Malanda B, Karuranga S, Unwin N, Colagiuri S, Guariguata L, Motala AA, Ogurtsova K (2019) Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: results from the International Diabetes Federation Diabetes Atlas, 9(th) edition. Diabetes Res Clin Pract 157:107843

[117] Schutgens F, Clevers H (2020) Human organoids: tools for understanding biology and treating diseases. Annu Rev Pathol 15:211–234

[118] Shang L, Hua H, Foo K, Martinez H, Watanabe K, Zimmer M, Kahler DJ, Freeby M, Chung W, LeDuc C (2014) beta-cell dysfunction due to increased ER stress in a stem cell model of Wolfram syndrome. Diabetes 63:923–933

[119] Sharon N, Vanderhooft J, Straubhaar J, Mueller J, Chawla R, Zhou Q, Engquist EN, Trapnell C, Gifford DK, Melton DA (2019) Wnt signaling separates the progenitor and endocrine compartments during pancreas development. Cell Rep 27(2281–2291):e2285

[120] Shi ZD, Lee K, Yang D, Amin S, Verma N, Li QV, Zhu Z, Soh CL, Kumar R, Evans T (2017) Genome editing in hPSCs reveals GATA6 haploinsufficiency and a genetic interaction with GATA4 in human pancreatic development. Cell Stem Cell 20(675–688): e676

[121] Shih HP, Wang A, Sander M (2013) Pancreas organogenesis: from lineage determination to morphogenesis. Annu Rev Cell Dev Biol 29:81–105

[122] Sneddon JB, Tang Q, Stock P, Bluestone JA, Roy S, Desai T, Hebrok M (2018) Stem cell therapies for treating diabetes: progress and remaining challenges. Cell Stem Cell 22:810–823

[123] Suckale J, Solimena M (2010) The insulin secretory granule as a signaling hub. Trends Endocrinol Metab 21:599–609

[124] Suzuki T, Itoh Y, Sakai Y, Saito A, Okuzaki D, Motooka D, Minami S, Kobayashi T, Yamamoto T, Okamoto T (2020) Generation of human bronchial organoids for SARS-CoV-2 research. bioRxiv

[125] Takahashi Y, Sekine K, Kin T, Takebe T, Taniguchi H (2018a) Selfcondensation culture enables vascularization of tissue fragments for efficient therapeutic transplantation. Cell Rep 23:1620–1629

[126] Takahashi Y, Takebe T, Taniguchi H (2018b) Methods for generating vascularized islet-like organoids via self-condensation. Curr Protoc Stem Cell Biol 45:e49

[127] Tao T, Wang Y, Chen W, Li Z, Su W, Guo Y, Deng P, Qin J (2019) Engineering human islet organoids from iPSCs using an organon-chip platform. Lab Chip 19:948–958

[128] Thorel F, Nepote V, Avril I, Kohno K, Desgraz R, Chera S, Herrera PL (2010) Conversion of adult pancreatic alpha-cells to beta-cells after extreme beta-cell loss. Nature 464:1149–1154

[129] Toyoda T, Mae S, Tanaka H, Kondo Y, Funato M, Hosokawa Y, Sudo T, Kawaguchi Y, Osafune K (2015) Cell aggregation optimizes the differentiation of human ESCs and iPSCs into pancreatic bud-like progenitor cells. Stem Cell Res 14:185–197

[130] Van de Casteele M, Leuckx G, Baeyens L, Cai Y, Yuchi Y, Coppens V, De Groef S, Eriksson M, Svensson C, Ahlgren U (2013) Neurogenin 3+ cells contribute to beta-cell neogenesis and proliferation in injured adult mouse pancreas. Cell Death Dis 4: e523

[131] Velazco-Cruz L, Song J, Maxwell KG, Goedegebuure MM, Augsornworawat P, Hogrebe NJ, Millman JR (2019) Acquisition of dynamic function in human stem cell-derived beta cells. Stem Cell Rep 12:351–365

[132] Velazco-Cruz L, Goedegebuure MM, Maxwell KG, Augsornworawat P, Hogrebe NJ, Millman JR (2020) SIX2 regulates human beta cell differentiation from stem cells and functional maturation in vitro. Cell Rep 31:107687

[133] Veres A, Faust AL, Bushnell HL, Engquist EN, Kenty JH, Harb G, Poh YC, Sintov E, Gurtler M, Pagliuca FW (2019) Charting cellular identity during human in vitro beta-cell differentiation. Nature 569:368–373

[134] Vethe H, Bjorlykke Y, Ghila LM, Paulo JA, Scholz H, Gygi SP, Chera S, Raeder H (2017) Probing the missing mature beta-cell proteomic landscape in differentiating patient iPSC-derived cells. Sci Rep 7:4780

[135] Viacyte (2014) A safety, tolerability, and efficacy study of VC-01^TM combination product in subjects with type I diabetes mellitus

[136] Viacyte (2016) One-year follow-up safety study in subjects previ-ously implanted with VC-01™

[137] Viacyte (2017a) A safety and tolerability study of VC-02 ^TM combination product in subjects with type 1 diabetes mellitus

[138] Viacyte (2017b) A safety, tolerability, and efficacy study of VC-02 ^TM combination product in subjects with type 1 diabetes mellitus and hypoglycemia unawareness

[139] Viacyte (2019) A study to evaluate safety, engraftment, and efficacy of VC-01 in subjects with T1 diabetes mellitus

[140] Wan J, Huang Y, Zhou P, Guo Y, Wu C, Zhu S, Wang Y, Wang L, Lu Y, Wang Z (2017) Culture of iPSCs derived pancreatic beta-like cells in vitro using decellularized pancreatic scaffolds: a preliminary trial. Biomed Res Int 2017:4276928

[141] Wang L, Zhan Y, Song E, Yu Y, Jiu Y, Du W, Lu J, Liu P, Xu P, Xu T (2011) HID-1 is a peripheral membrane protein primarily associated with the medial- and trans- Golgi apparatus. Protein Cell 2:74–85

[142] Wang W, Jin S, Ye K (2017) Development of islet organoids from H9 human embryonic stem cells in biomimetic 3D scaffolds. Stem Cells Dev 26:394–404

[143] Wang D, Wang J, Bai L, Pan H, Feng H, Clevers H, Zeng YA (2020) Long-term expansion of pancreatic islet organoids from resident procr(+) progenitors. Cell 180(1198–1211):e1119

[144] Wei P, Li L, Qi H, Zhou HX, Deng CY, Li FR (2012) Reversible immortalization of Nestin-positive precursor cells from pancreas and differentiation into insulin-secreting cells. Biochem Biophys Res Commun 418:330–335

[145] Wei Z, Yoshihara E, He N, Hah N, Fan W, Pinto AFM, Huddy T, Wang Y, Ross B, Estepa G (2018) Vitamin D switches BAF complexes to protect beta cells. Cell 173(1135–1149):e1115

[146] Weng C, Xi J, Li H, Cui J, Gu A, Lai S, Leskov K, Ke L, Jin F, Li Y (2020) Single-cell lineage analysis reveals extensive multimodal transcriptional control during directed beta-cell differentiation. Nat Metab 2:1443–1458

[147] Wesolowska-Andersen A, Jensen RR, Alcántara MP, Beer NL, Duff C, Nylander V, Gosden M, Witty L, Bowden R, McCarthy MI (2020) Analysis of differentiation protocols defines a common pancreatic progenitor molecular signature and guides refinement of endocrine differentiation. Stem Cell Rep 14:138–153

[148] Wu Y, Chang T, Long Y, Huang H, Kandeel F, Yee JK (2019) Using gene editing to establish a safeguard system for pluripotent stemcell-based therapies. iScience 22:409–422

[149] Yabe SG, Iwasaki N, Yasuda K, Hamazaki TS, Konno M, Fukuda S, Takeda F, Kasuga M, Okochi H (2015) Establishment of maturityonset diabetes of the young-induced pluripotent stem cells from a Japanese patient. J Diabetes Investig 6:543–547

[150] Yang L, Han Y, Nilsson-Payant BE, Gupta V, Wang P, Duan X, Tang X, Zhu J, Zhao Z, Jaffre F (2020) A human pluripotent stem cell-based platform to study SARS-CoV-2 tropism and model virus infection in human cells and organoids. Cell Stem Cell 27 (125–136):e127

[151] Yoshihara E, Wei Z, Lin CS, Fang S, Ahmadian M, Kida Y, Tseng T, Dai Y, Yu RT, Liddle C (2016) ERRgamma is required for the metabolic maturation of therapeutically functional glucose-responsive beta cells. Cell Metab 23:622–634

[152] Yoshihara E, O’Connor C, Gasser E, Wei Z, Oh TG, Tseng TW, Wang D, Cayabyab F, Dai Y, Yu RT (2020) Immune-evasive human islet-like organoids ameliorate diabetes. Nature 586:606–611

[153] Youn DY, Xiaoli AM, Pessin JE, Yang F (2016) Regulation of metabolism by the Mediator complex. Biophys Rep 2:69–77

[154] Youngblood RL, Sampson JP, Lebioda KR, Shea LD (2019) Microporous scaffolds support assembly and differentiation of pancreatic progenitors into beta-cell clusters. Acta Biomater 96:111–122

[155] Yu XX, Xu CR (2020) Understanding generation and regeneration of pancreatic beta cells from a single-cell perspective. Development147

[156] Zeng H, Guo M, Zhou T, Tan L, Chong CN, Zhang T, Dong X, Xiang JZ, Yu AS, Yue L (2016) An isogenic human ESC platform for functional evaluation of genome-wide-association-study-identified diabetes genes and drug discovery. Cell Stem Cell 19:326–340

[157] Zhang X, McGrath PS, Salomone J, Rahal M, McCauley HA, Schweitzer J, Kovall R, Gebelein B, Wells JM (2019) A comprehensive structure-function study of neurogenin3 disease-causing alleles during human pancreas and intestinal organoid development. Dev Cell 50(367–380):e367

[158] Zhou T, Kim TW, Chong CN, Tan L, Amin S, Sadat Badieyan Z, Mukherjee S, Ghazizadeh Z, Zeng H, Guo M (2018) A hPSC-based platform to discover gene-environment interactions that impact human beta-cell and dopamine neuron survival. Nat Commun 9:4815

[159] Zhou X, Nair GG, Russ HA, Belair CD, Li ML, Shveygert M, Hebrok M, Blelloch R (2020) LIN28B impairs the transition of hESC- derived beta cells from the juvenile to adult state. Stem Cell Rep 14:9–20

[160] Zhu P, Fan Z (2018) Cancer stem cells and tumorigenesis. Biophys Rep 4:178–188

[161] Zhu Z, Li QV, Lee K, Rosen BP, Gonzalez F, Soh CL, Huangfu D (2016) Genome editing of lineage determinants in human pluripotent stem cells reveals mechanisms of pancreatic development and diabetes. Cell Stem Cell 18:755–768

[162] Zhu L, She ZG, Cheng X, Qin JJ, Zhang XJ, Cai J, Lei F, Wang H, Xie J, Wang W (2020) Association of blood glucose control and outcomes in patients with COVID-19 and pre-existing type 2 diabetes. Cell Metab 31(1068–1077):e1063

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