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Diabetologia

Erschienen in:

01.04.2005 | Article

Efficient restricted gene expression in beta cells by lentivirus-mediated gene transfer into pancreatic stem/progenitor cells

verfasst von: M. Castaing, A. Guerci, J. Mallet, P. Czernichow, P. Ravassard, R. Scharfmann

Erschienen in: Diabetologia | Ausgabe 4/2005

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Abstract

Aims/hypothesis

Gene transfer into pancreatic beta cells, which produce and secrete insulin, is a promising strategy to protect such cells against autoimmune destruction and also to generate beta cells in mass, thereby providing a novel therapeutic approach to treat diabetic patients. Until recently, exogenous DNA has been directly transferred into mature beta cells with various levels of success. We investigated whether exogenous DNA could be stably transferred into pancreatic stem/progenitor cells, which would subsequently differentiate into mature beta cells expressing the transgene.

Methods

We designed transplantation and tissue culture procedures to obtain ex vivo models of pancreatic development. We next constructed recombinant lentiviruses expressing enhanced green fluorescent protein (eGFP) under the control of either the rat insulin promoter or a ubiquitous promoter, and performed viral infection of rat embryonic pancreatic tissue.

Results

Embryonic pancreas infected with recombinant lentiviruses resulted in endocrine cell differentiation and restricted cell type expression of the transgene according to the specificity of the promoter used in the viral construct. We next demonstrated that the efficiency of infection could be further improved upon infection of embryonic pancreatic epithelia, followed by their in vitro culture, using conditions that favour endocrine cell differentiation. Under these conditions, endocrine stem/progenitor cells expressing neurogenin 3 are efficiently transduced by recombinant lentiviral vectors. Moreover, when eGFP was placed under the control of the insulin promoter, 70.4% of the developed beta cells were eGFP-expressing cells. All of the eGFP-positive cells were insulin-producing cells.

Conclusions/interpretation

We have demonstrated that mature rat pancreatic beta cells can be stably modified by infecting pancreatic stem/progenitor cells that undergo endocrine differentiation.

Shapiro AM, Lakey JR, Ryan EA et al (2000) Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med 343:230–238CrossRefPubMed

Halban PA, Kahn SE, Lernmark A, Rhodes CJ (2001) Gene and cell-replacement therapy in the treatment of type 1 diabetes: how high must the standards be set? Diabetes 50:2181–2191

Leibowitz G, Beattie GM, Kafri T et al (1999) Gene transfer to human pancreatic endocrine cells using viral vectors. Diabetes 48:745–753

Csete ME, Benhamou PY, Drazan KE et al (1995) Efficient gene transfer to pancreatic islets mediated by adenoviral vectors. Transplantation 59:263–268

Becker TC, BeltrandelRio H, Noel RJ, Johnson JH, Newgard CB (1994) Overexpression of hexokinase I in isolated islets of Langerhans via recombinant adenovirus. Enhancement of glucose metabolism and insulin secretion at basal but not stimulatory glucose levels. J Biol Chem 269:21234–21238

Verma IM, Somia N (1997) Gene therapy—promises, problems and prospects. Nature 389:239–242

Kapturczak M, Zolotukhin S, Cross J et al (2002) Transduction of human and mouse pancreatic islet cells using a bicistronic recombinant adeno-associated viral vector. Mol Ther 5:154–160

Flotte T, Agarwal A, Wang J et al (2001) Efficient ex vivo transduction of pancreatic islet cells with recombinant adeno-associated virus vectors. Diabetes 50:515–520

Ju Q, Edelstein D, Brendel MD et al (1998) Transduction of non-dividing adult human pancreatic beta cells by an integrating lentiviral vector. Diabetologia 41:736–739

10.

Gallichan WS, Kafri T, Krahl T, Verma IM, Sarvetnick N (1998) Lentivirus-mediated transduction of islet grafts with interleukin 4 results in sustained gene expression and protection from insulitis. Hum Gene Ther 9:2717–2726

11.

Salmon P, Oberholzer J, Occhiodoro T, Morel P, Lou J, Trono D (2000) Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes. Mol Ther 2:404–414

12.

Uchida N, Buck DW, He D et al (2000) Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci U S A 97:14720–14725

13.

Case SS, Price MA, Jordan CT et al (1999) Stable transduction of quiescent CD34(+)CD38(-) human hematopoietic cells by HIV-1-based lentiviral vectors. Proc Natl Acad Sci U S A 96:2988–2993

14.

Miyoshi H, Smith KA, Mosier DE, Verma IM, Torbett BE (1999) Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors. Science 283:682–686

15.

Evans JT, Kelly PF, O’Neill E, Garcia JV (1999) Human cord blood CD34+CD38-cell transduction via lentivirus-based gene transfer vectors. Hum Gene Ther 10:1479–1489

16.

Pawliuk R, Westerman KA, Fabry ME et al (2001) Correction of sickle cell disease in transgenic mouse models by gene therapy. Science 294:2368–2371

17.

Buchet D, Serguera C, Zennou V, Charneau P, Mallet J (2002) Long-term expression of beta-glucuronidase by genetically modified human neural progenitor cells grafted into the mouse central nervous system. Mol Cell Neurosci 19:389–401

18.

Flax JD, Aurora S, Yang C et al (1998) Engraftable human neural stem cells respond to developmental cues, replace neurons, and express foreign genes. Nat Biotechnol 16:1033–1039CrossRefPubMed

19.

Englund U, Ericson C, Rosenblad C et al (2000) The use of a recombinant lentiviral vector for ex vivo gene transfer into the rat CNS. NeuroReport 11:3973–3977

20.

Zennou V, Petit C, Guetard D, Nerhbass U, Montagnier L, Charneau P (2000) HIV-1 genome nuclear import is mediated by a central DNA flap. Cell 101:173–185

21.

Sanvito F, Nichols A, Herrera P et al (1995) TGF-β1 overexpression in murine pancreas induces chronic pancreatitis and, together with TNF-α, triggers insulin-dependent diabetes. Biochem Biophys Res Commun 217:1279–1286CrossRefPubMed

22.

Zufferey R, Nagy D, Mandel RJ, Naldini L, Trono D (1997) Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 15:871–875

23.

Yee JK, Miyanohara A, LaPorte P, Bouic K, Burns JC, Friedmann T (1994) A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes. Proc Natl Acad Sci U S A 91:9564–9568

24.

Miyazaki J, Araki K, Yamato E et al (1990) Establishment of a pancreatic beta cell line that retains glucose-inducible insulin secretion: special reference to expression of glucose transporter isoforms. Endocrinology 127:126–132

25.

Gregory LG, Harbottle RP, Lawrence L, Knapton HJ, Themis M, Coutelle C (2003) Enhancement of adenovirus-mediated gene transfer to the airways by DEAE dextran and sodium caprate in vivo. Mol Ther 7:19–26

26.

Gittes G, Galante P, Hanahan D, Rutter W, Debas H (1996) Lineage specific morphogenesis in the developing pancreas: role of mesenchymal factors. Development 122:439–447PubMed

27.

Miralles F, Czernichow P, Scharfmann R (1998) Follistatin regulates the relative proportions of endocrine versus exocrine tissue during pancreatic development. Development 125:1017–1024

28.

Rachdi L, Marie JC, Scharfmann R (2003) Role for VPAC2 receptor-mediated signals in pancreas development. Diabetes 52:85–92

29.

Castaing M, Peault B, Basmaciogullari A, Casal I, Czernichow P, Scharfmann R (2001) Blood glucose normalization upon transplantation of human embryonic pancreas into beta-cell-deficient SCID mice. Diabetologia 44:2066–2076CrossRef

30.

Thomas M, Northrup SR, Hornsby PJ (1997) Adrenocortical tissue formed by transplantation of normal clones of bovine adrenocortical cells in scid mice replaces the essential functions of the animals’ adrenal glands. Nat Med 3:978–983

31.

Cras-Meneur C, Elghazi L, Czernichow P, Scharfmann R (2001) Epidermal growth factor increases undifferentiated pancreatic embryonic cells in vitro: a balance between proliferation and differentiation. Diabetes 50:1571–1579PubMed

32.

Ravassard P, Chatail F, Mallet J, Icard-Liepkalns C (1997) Relax, a novel rat bHLH transcriptional regulator transiently expressed in the ventricular proliferating zone of the developing central nervous system. J Neurosci Res 48:146–158

33.

Herzog E, Bellenchi GC, Gras C et al (2001) The existence of a second vesicular glutamate transporter specifies subpopulations of glutamatergic neurons. J Neurosci 21:RC181

34.

Rall LB, Pictet RL, Williams RH, Rutter WJ (1973) Early differentiation of glucagon-producing cells in embryonic pancreas: a possible developmental role for glucagon. Proc Natl Acad Sci U S A 70:3478–3482

35.

Miralles F, Serup P, Cluzeaud F, Vandewalle A, Czernichow P, Scharfmann R (1999) Characterization of beta cells developed in vitro from rat embryonic pancreatic epithelium. Dev Dyn 214:116–126

36.

Gradwohl G, Dierich A, LeMeur M, Guillemot F (2000) Neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas. Proc Natl Acad Sci U S A 97:1607–1611

37.

Gu G, Dubauskaite J, Melton DA (2002) Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors. Development 129:2447–2457

38.

Demeterco C, Levine F (2001) Gene therapy for diabetes. Front Biosci 6:D175–D191

39.

De la Tour D, Halvorsen T, Demeterco C et al (2001) Beta-cell differentiation from a human pancreatic cell line in vitro and in vivo. Mol Endocrinol 15:476–483

40.

Hoffmann MW, Allison J, Miller JF (1992) Tolerance induction by thymic medullary epithelium. Proc Natl Acad Sci U S A 89:2526–2530

41.

Hayek A, Beattie GM (1997) Experimental transplantation of human fetal and adult pancreatic islets. J Clin Endocrinol Metab 82:2471–2475

42.

Jensen J, Heller RS, Funder-Nielsen T et al (2000) Independent development of pancreatic alpha- and beta-cells from neurogenin3-expressing precursors: a role for the notch pathway in repression of premature differentiation. Diabetes 49:163–176PubMed

43.

Herrera PL (2000) Adult insulin- and glucagon-producing cells differentiate from two independent cell lineages. Development 127:2317–2322PubMed

44.

Gu G, Brown JR, Melton DA (2003) Direct lineage tracing reveals the ontogeny of pancreatic cell fates during mouse embryogenesis. Mech Dev 120:35–43

45.

Apelqvist A, Li H, Sommer L et al (1999) Notch signalling controls pancreatic cell differentiation. Nature 400:877–881CrossRefPubMed

46.

Lee JC, Smith SB, Watada H et al (2001) Regulation of the pancreatic pro-endocrine gene neurogenin3. Diabetes 50:928–936

47.

Polak M, Bouchareb-Banaei L, Scharfmann R, Czernichow P (2000) Early pattern of differentiation in the human pancreas. Diabetes 49:225–232

48.

Abbas-Terki T, Blanco-Bose W, Deglon N, Pralong W, Aebischer P (2002) Lentiviral-mediated RNA interference. Hum Gene Ther 13:2197–2201

49.

Rubinson DA, Dillon CP, Kwiatkowski AV et al (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat Genet 33:401–406CrossRefPubMed

50.

Stewart SA, Dykxhoorn DM, Palliser D et al (2003) Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9:493–501

51.

Efrat S, Linde S, Kofod H et al (1988) β Cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc Natl Acad Sci U S A 85:9037–9041

Titel: Efficient restricted gene expression in beta cells by lentivirus-mediated gene transfer into pancreatic stem/progenitor cells
verfasst von: M. Castaing
A. Guerci
J. Mallet
P. Czernichow
P. Ravassard
R. Scharfmann
Publikationsdatum: 01.04.2005
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 4/2005
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-005-1694-6

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