Sustained secretion of human growth hormone from TheraCyte devices encapsulated with PiggyBac-engineered retinal pigment epithelium cells

dc.creatorCECCHI, CLAUDIA R.
dc.creatorALSING, SIDSEL
dc.creatorJESUS, GUSTAVO P.P.
dc.creatorZACARIAS, ENIO A.
dc.creatorKJAER, LISBETH
dc.creatorCLEMENT, MICHELLE S.
dc.creatorKUMAGAI-BRAESCH, MAKIKO
dc.creatorCORYDON, THOMAS J.
dc.creatorBARTOLINI, PAOLO
dc.creatorPERONI, CIBELE N.
dc.creatorAAGAARD, LARS
dc.date2023
dc.date2023-06-26T13:50:55Z
dc.date2023-06-26T13:50:55Z
dc.date.accessioned2023-09-28T14:26:07Z
dc.date.available2023-09-28T14:26:07Z
dc.identifier0040-8166
dc.identifierhttp://repositorio.ipen.br/handle/123456789/34084
dc.identifier82
dc.identifier10.1016/j.tice.2023.102095
dc.identifier51.4
dc.identifier27
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/9004292
dc.descriptionGrowth hormone (GH) deficiency is characterized by impaired growth and development, and is currently treated by repeated administration of recombinant human GH (hGH). Encapsulated cell therapy (ECT) may offer a less demanding treatment-strategy for long-term production and release of GH into circulation. We used PiggyBac-based (PB) transposon delivery for engineering retinal pigment epithelial cells (ARPE-19), and tested a series of viral and non-viral promoters as well as codon-optimization to enhance transgene expression. Engineered cells were loaded into TheraCyte macrocapsules and secretion was followed in vitro and in vivo. The cytomegalovirus (CMV) promoter supports strong and persistent transgene expression, and we achieved clonal cell lines secreting over 6 ??g hGH/106 cells/day. Codon-optimization of the hGH gene did not improve secretion. ARPE-19 cells endured encapsulation in TheraCyte devices, and resulted in steady hormone release for at least 60 days in vitro. A short-term pilot experiment in immunodeficient SCID mice demonstrated low systemic levels of hGH from a single 40 ??L capsule implanted subcutaneously. No significant increase in weight increase or systemic hGH was detected after 23 days in the GH-deficient lit/SCID mouse model using 4.5 ??L capsules loaded with the highest secreting clone of ARPE-19 cells. Our results demonstrate that PB-mediated engineering of ARPE-19 is an efficient way to generate hormone secreting cell lines compatible with macroencapsulation, and our CMV-driven expression cassette allows for identification of clones with high level and long-term secretory activity without addition of insulator elements. Our results pave the way for further in vivo studies of encapsulated cell therapy.
dc.descriptionFunda????o de Amparo ?? Pesquisa do Estado de S??o Paulo (FAPESP)
dc.descriptionFrimodt-Heineke Foundation
dc.descriptionFAPESP: 13/03747-0; 14/04277-0; 14/07380-6; 14/18242-3; 14/19757-7
dc.descriptionLundbeckfonden: R126- 2012-12456
dc.format1-13
dc.relationTissue and Cell
dc.rightsopenAccess
dc.subjecthormones
dc.subjectsth
dc.subjectepithelium
dc.subjectpigments
dc.subjecttherapy
dc.titleSustained secretion of human growth hormone from TheraCyte devices encapsulated with PiggyBac-engineered retinal pigment epithelium cells
dc.typeArtigo de peri??dico
dc.coverageI


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