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Abstract: TH-OR136

Therapeutic Effects of Genome-Engineered Angiogenic or Anti-Inflammatory Factor Secreting-Mesenchymal Stem Cells in Mice with AKI

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 502 Development, Stem Cells, and Regenerative Medicine: Clinical

Authors

  • Park, Hye-Jeong, Kyungpook National University, Taegu, Korea (the Republic of)
  • Kong, Min Jung, Kyungpook National University, Taegu, Korea (the Republic of)
  • Jang, Hyo-Ju, Kyungpook National University, Taegu, Korea (the Republic of)
  • Park, Eui-Jung, Kyungpook National University, Taegu, Korea (the Republic of)
  • Choi, Hyo-Jung, Kyungpook National University, Taegu, Korea (the Republic of)
  • Jung, Hyun Jun, NHLBI/NIH, Bethesda, Maryland, United States
  • Park, Kwon Moo, Kyungpook National University, Taegu, Korea (the Republic of)
  • Kwon, Tae-Hwan, Kyungpook National University, Taegu, Korea (the Republic of)
Background

Stem cell therapy has been proposed as a potential therapeutic strategy for acute kidney injury (AKI). By exploiting genome editing technology, we generated mesenchymal stem cells (MSCs) secreting angiogenic factor (angiopoietin 1: ANG1) or anti-inflammatory factor (erythropoietin: EPO) for therapeutic application in AKI.

Methods

To integrate each gene cassette into the safe harbor locus, AAVS1, of the human umbilical cord-derived MSCs (hUC-MSCs) chromosome, AAVS1-targeting Zinc Finger Nuclease (ZFN) system was exploited. For the application of stem cell therapy in vivo, a scaffold-free cell sheet system was established using a temperature-responsive polymer. AKI in C57BL/6 mice was established by bilateral renal pedicles occlusion for 30 min and monitored for additional 7 days after the clamps were removed.

Results

Junction PCR analysis demonstrated the ZFN-aided gene integration in hUC-MSCs. Flow cytometry and osteogenic and adipogenic differentiation assay revealed that genome editing did not affect the stemness. Protein measurement in conditioned media by ELISA and immunoblotting confirmed the production and secretion of each integrated gene product (ANG1 and EPO proteins). The in vitro angiogenic function of hUC-MSCs secreting ANG1 was further revealed by the increased cell migration and expression of MMP-9 and Tie-2 mRNAs in co-cultured HUVEC. As a control experiment, a mono-layered sheet of hUC-MSCs was applied to the mice kidney surface in vivo. Immunohistochemistry with anti-human nuclei antibody at 1 or 2 weeks after the application demonstrated that hUC-MSCs on the mice kidney surface were intact and specifically labeled. In mice subjected to bilateral renal ischemia and reperfusion, cell sheets of ANG1- or EPO-secreting hUC-MSCs applied to the kidney surface significantly ameliorated renal functional deterioration with lower BUN and creatinine levels, compared with sham control mice as well as mice treated with sheets of GFP-expressing hUC-MSCs. Moreover, treatment of EPO-secreting hUC-MSCs resulted in higher Hct levels.

Conclusion

A novel cell therapy of hUC-MSCs secreting ANG1 or EPO provides the therapeutic effects against AKI in vivo.

Funding

  • Government Support - Non-U.S.