Alexandra Norrick1, Jasmina Esterlechner1, Elke Niebergall-Roth1, Markus H Frank2,3,4,5, Mark A Kluth6,7, Ulf Dehio8, Samar Sadeghi1, Hannes M Schröder8, Seda Ballikaya1, Nicole Stemler1, Christoph Ganss1,8, Kathrin Dieter8, Ann-Kathrin Dachtler8, Patrick Merz9,10, Saadettin Sel10, James Chodosh11, Claus Cursiefen12,13, Natasha Y Frank14,15,2,3, Gerd U Auffarth9,10, Bruce Ksander16. 1. TICEBA GmbH, Im Neuenheimer Feld 517, 69120, Heidelberg, Germany. 2. Transplant Research Program, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA. 3. Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA. 4. Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. 5. School of Medical and Health Sciences, Edith Cowan University, Perth, Western Australia, Australia. 6. TICEBA GmbH, Im Neuenheimer Feld 517, 69120, Heidelberg, Germany. andreas.kluth@ticeba.com. 7. RHEACELL GmbH & Co. KG, Im Neuenheimer Feld 517, Heidelberg, 69120, Germany. andreas.kluth@ticeba.com. 8. RHEACELL GmbH & Co. KG, Im Neuenheimer Feld 517, Heidelberg, 69120, Germany. 9. Department of Ophthalmology, Lions Eye Bank, University of Heidelberg, Heidelberg, Germany. 10. Department of Ophthalmology, University Hospital Heidelberg, Heidelberg, Germany. 11. Department of Ophthalmology, Massachusetts Eye & Ear, Harvard Medical School, Boston, MA, USA. 12. Department of Ophthalmology, University Hospital Cologne, Cologne, Germany. 13. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany. 14. Department of Medicine, VA Boston Healthcare System, Boston, MA, USA. 15. Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. 16. The Schepens Eye Research Institute, Massachusetts Eye & Ear, Harvard Medical School, Boston, MA, USA.
Abstract
BACKGROUND: While therapeutic success of the limbal tissue or cell transplantation to treat severe cases of limbal stem cell (LSC) deficiency (LSCD) strongly depends on the percentage of LSCs within the transplanted cells, prospective LSC enrichment has been hampered by the intranuclear localization of the previously reported LSC marker p63. The recent identification of the ATP-binding cassette transporter ABCB5 as a plasma membrane-spanning marker of LSCs that are capable of restoring the cornea and the development of an antibody directed against an extracellular loop of the ABCB5 molecule stimulated us to develop a novel treatment strategy based on the utilization of in vitro expanded allogeneic ABCB5+ LSCs derived from human cadaveric limbal tissue. METHODS: We developed and validated a Good Manufacturing Practice- and European Pharmacopeia-conform production and quality-control process, by which ABCB5+ LSCs are derived from human corneal rims, expanded ex vivo, isolated as homogenous cell population, and manufactured as an advanced-therapy medicinal product (ATMP). This product was tested in a preclinical study program investigating the cells' engraftment potential, biodistribution behavior, and safety. RESULTS: ABCB5+ LSCs were reliably expanded and manufactured as an ATMP that contains comparably high percentages of cells expressing transcription factors critical for LSC stemness maintenance (p63) and corneal epithelial differentiation (PAX6). Preclinical studies confirmed local engraftment potential of the cells and gave no signals of toxicity and tumorgenicity. These findings were sufficient for the product to be approved by the German Paul Ehrlich Institute and the U.S. Food & Drug Administration to be tested in an international multicenter phase I/IIa clinical trial (NCT03549299) to evaluate the safety and therapeutic efficacy in patients with LSCD. CONCLUSION: Building upon these data in conjunction with the previously shown cornea-restoring capacity of human ABCB5+ LSCs in animal models of LSCD, we provide an advanced allogeneic LSC-based treatment strategy that shows promise for replenishment of the patient's LSC pool, recreation of a functional barrier against invading conjunctival cells and restoration of a transparent, avascular cornea.
BACKGROUND: While therapeutic success of the limbal tissue or cell transplantation to treat severe cases of limbal stem cell (LSC) deficiency (LSCD) strongly depends on the percentage of LSCs within the transplanted cells, prospective LSC enrichment has been hampered by the intranuclear localization of the previously reported LSC marker p63. The recent identification of the ATP-binding cassette transporter ABCB5 as a plasma membrane-spanning marker of LSCs that are capable of restoring the cornea and the development of an antibody directed against an extracellular loop of the ABCB5 molecule stimulated us to develop a novel treatment strategy based on the utilization of in vitro expanded allogeneic ABCB5+ LSCs derived from human cadaveric limbal tissue. METHODS: We developed and validated a Good Manufacturing Practice- and European Pharmacopeia-conform production and quality-control process, by which ABCB5+ LSCs are derived from human corneal rims, expanded ex vivo, isolated as homogenous cell population, and manufactured as an advanced-therapy medicinal product (ATMP). This product was tested in a preclinical study program investigating the cells' engraftment potential, biodistribution behavior, and safety. RESULTS: ABCB5+ LSCs were reliably expanded and manufactured as an ATMP that contains comparably high percentages of cells expressing transcription factors critical for LSC stemness maintenance (p63) and corneal epithelial differentiation (PAX6). Preclinical studies confirmed local engraftment potential of the cells and gave no signals of toxicity and tumorgenicity. These findings were sufficient for the product to be approved by the German Paul Ehrlich Institute and the U.S. Food & Drug Administration to be tested in an international multicenter phase I/IIa clinical trial (NCT03549299) to evaluate the safety and therapeutic efficacy in patients with LSCD. CONCLUSION: Building upon these data in conjunction with the previously shown cornea-restoring capacity of human ABCB5+ LSCs in animal models of LSCD, we provide an advanced allogeneic LSC-based treatment strategy that shows promise for replenishment of the patient's LSC pool, recreation of a functional barrier against invading conjunctival cells and restoration of a transparent, avascular cornea.
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