Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells
Direct nuclear reprogramming has the potential to enable the development of β cell replacement therapies for diabetes that do not require the use of progenitor/stem cell populations. However, despite their promise, current approaches to β cell‐directed reprogramming rely heavily on the use of viral...
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley-VCH
2023-10-01
|
| Series: | Advanced NanoBiomed Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/anbr.202200173 |
| _version_ | 1827793245842702336 |
|---|---|
| author | Lilibeth Ortega-Pineda Maria Angelica Rincon-Benavides Tatiana Z. Cuellar-Gaviria Mia Kordowski Elizabeth Guilfoyle Amrita Lakshmi Anaparthi Luke R. Lemmerman William Lawrence Jill L. Buss Binbin Deng Britani N. Blackstone Ana Salazar-Puerta David W. McComb Heather Powell Daniel Gallego-Perez Natalia Higuita-Castro |
| author_facet | Lilibeth Ortega-Pineda Maria Angelica Rincon-Benavides Tatiana Z. Cuellar-Gaviria Mia Kordowski Elizabeth Guilfoyle Amrita Lakshmi Anaparthi Luke R. Lemmerman William Lawrence Jill L. Buss Binbin Deng Britani N. Blackstone Ana Salazar-Puerta David W. McComb Heather Powell Daniel Gallego-Perez Natalia Higuita-Castro |
| author_sort | Lilibeth Ortega-Pineda |
| collection | DOAJ |
| description | Direct nuclear reprogramming has the potential to enable the development of β cell replacement therapies for diabetes that do not require the use of progenitor/stem cell populations. However, despite their promise, current approaches to β cell‐directed reprogramming rely heavily on the use of viral vectors. Herein, the use of extracellular vesicles (EVs) derived from human dermal fibroblasts (HDFs) is explored as novel nonviral carriers of endocrine cell‐patterning transcription factors, to transfect and transdifferentiate pancreatic ductal epithelial cells (PDCs) into hormone‐expressing cells. Electrotransfection of HDFs with expression plasmids for Pdx1, Ngn3, and MafA (PNM) leads to the release of EVs loaded with PNM at the gene, mRNA, and protein levels. Exposing PDC cultures to PNM‐loaded EVs leads to successful transfection and increases PNM expression in PDCs, which ultimately result in endocrine cell‐directed conversions based on the expression of insulin/c‐peptide, glucagon, and glucose transporter 2 (Glut2). These findings are further corroborated in vivo in a mouse model following intraductal injection of PNM‐ versus sham‐loaded EVs. Collectively, these findings suggest that dermal fibroblast‐derived EVs can potentially serve as a powerful platform technology for the development and deployment of nonviral reprogramming‐based cell therapies for insulin‐dependent diabetes. |
| first_indexed | 2024-03-11T18:15:46Z |
| format | Article |
| id | doaj.art-032b7c11666d41b9a6180c90fb305ad6 |
| institution | Directory Open Access Journal |
| issn | 2699-9307 |
| language | English |
| last_indexed | 2024-03-11T18:15:46Z |
| publishDate | 2023-10-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Advanced NanoBiomed Research |
| spelling | doaj.art-032b7c11666d41b9a6180c90fb305ad62023-10-16T07:03:54ZengWiley-VCHAdvanced NanoBiomed Research2699-93072023-10-01310n/an/a10.1002/anbr.202200173Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal CellsLilibeth Ortega-Pineda0Maria Angelica Rincon-Benavides1Tatiana Z. Cuellar-Gaviria2Mia Kordowski3Elizabeth Guilfoyle4Amrita Lakshmi Anaparthi5Luke R. Lemmerman6William Lawrence7Jill L. Buss8Binbin Deng9Britani N. Blackstone10Ana Salazar-Puerta11David W. McComb12Heather Powell13Daniel Gallego-Perez14Natalia Higuita-Castro15Department of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USABiophysics -Graduate Program The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USABiomedical Science Graduate Program The Ohio State University Columbus OH 43210 USADepartment of Hematology and the Bloomfield Center for Leukemia Outcomes Research The Ohio State University Columbus OH 43210 USACenter for Electron Microscopy and Analysis (CEMAS) The Ohio State University Columbus OH 43212 USADepartment of Materials Science and Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USACenter for Electron Microscopy and Analysis (CEMAS) The Ohio State University Columbus OH 43212 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADepartment of Biomedical Engineering The Ohio State University Columbus OH 43210 USADirect nuclear reprogramming has the potential to enable the development of β cell replacement therapies for diabetes that do not require the use of progenitor/stem cell populations. However, despite their promise, current approaches to β cell‐directed reprogramming rely heavily on the use of viral vectors. Herein, the use of extracellular vesicles (EVs) derived from human dermal fibroblasts (HDFs) is explored as novel nonviral carriers of endocrine cell‐patterning transcription factors, to transfect and transdifferentiate pancreatic ductal epithelial cells (PDCs) into hormone‐expressing cells. Electrotransfection of HDFs with expression plasmids for Pdx1, Ngn3, and MafA (PNM) leads to the release of EVs loaded with PNM at the gene, mRNA, and protein levels. Exposing PDC cultures to PNM‐loaded EVs leads to successful transfection and increases PNM expression in PDCs, which ultimately result in endocrine cell‐directed conversions based on the expression of insulin/c‐peptide, glucagon, and glucose transporter 2 (Glut2). These findings are further corroborated in vivo in a mouse model following intraductal injection of PNM‐ versus sham‐loaded EVs. Collectively, these findings suggest that dermal fibroblast‐derived EVs can potentially serve as a powerful platform technology for the development and deployment of nonviral reprogramming‐based cell therapies for insulin‐dependent diabetes.https://doi.org/10.1002/anbr.202200173cell reprogrammingdiabetesengineered EVsnonviral gene deliveryβ cell replacement therapies |
| spellingShingle | Lilibeth Ortega-Pineda Maria Angelica Rincon-Benavides Tatiana Z. Cuellar-Gaviria Mia Kordowski Elizabeth Guilfoyle Amrita Lakshmi Anaparthi Luke R. Lemmerman William Lawrence Jill L. Buss Binbin Deng Britani N. Blackstone Ana Salazar-Puerta David W. McComb Heather Powell Daniel Gallego-Perez Natalia Higuita-Castro Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells Advanced NanoBiomed Research cell reprogramming diabetes engineered EVs nonviral gene delivery β cell replacement therapies |
| title | Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells |
| title_full | Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells |
| title_fullStr | Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells |
| title_full_unstemmed | Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells |
| title_short | Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells |
| title_sort | engineered extracellular vesicles from human skin cells induce pro β cell conversions in pancreatic ductal cells |
| topic | cell reprogramming diabetes engineered EVs nonviral gene delivery β cell replacement therapies |
| url | https://doi.org/10.1002/anbr.202200173 |
| work_keys_str_mv | AT lilibethortegapineda engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT mariaangelicarinconbenavides engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT tatianazcuellargaviria engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT miakordowski engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT elizabethguilfoyle engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT amritalakshmianaparthi engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT lukerlemmerman engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT williamlawrence engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT jilllbuss engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT binbindeng engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT britaninblackstone engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT anasalazarpuerta engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT davidwmccomb engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT heatherpowell engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT danielgallegoperez engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells AT nataliahiguitacastro engineeredextracellularvesiclesfromhumanskincellsinduceprobcellconversionsinpancreaticductalcells |