Project Details
Projekt
Development of a biohybrid lung - Preventing rejection of allogenic endothelial cells by HLA-class I silencing
Subject Area
Cardiac and Vascular Surgery
Term
from 2012 to 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 221323735
Due to the fact that there is currently no implantable long-term assist device for end-stage lung disease, lung transplantation remains the only curative option at present. The aim of the proposed project is the development of an implantable bioartifical lung as a potential long-term alternative to transplantation. Gas exchange membranes, which are used in the extracorporeal membrane oxygenators, will be endothelialized to improve hemocompatability. Patients for whom the bioartificial lung could be considered as a treatment option will typically not have enough autologous endothelial cells (EC) for cell seeding, rendering the application of allogenous EC necessary. This project will therefore focus on the analysis of HLA-silencing. By means of so-called RNAi-technology, the surface characteristics of the human leukocyte antigen (HLA), which are responsible for transplant rejection, will be suppressed, and therefore the silenced allogenous EC will not be detected by the immune system. The application of these silenced EC in the in vitro development of a confluent endothelial monolayer on the gas exchange membranes will be investigated. In addition to the optimization of the HLA-silencing, the suitability of silenced EC to endothelialize the gas exchange membranes will be analysed. For this purpose, analyses under static and dynamic conditions in combination with different surface modifications of the membranes, as well as the verification of the respective gas exchange capabilities, will be conducted. Functional analyses will include the investigation of the effectiveness of the HLA-silencing (hemocompatibility assays, analysis of cytotoxic T- and NK cell effects, adherence and migration of immunological cells) and the identification of the so-called off-target effects. In preparation for the in vivo application in a small animal model (rat), the isolation of EC, the establishment of endothelial specific primers and antibodies for PCR and FACS-analysis and the HLA-silencing of rat-specific EC, as well as the establishment of a suitable bioreactor, constitute the focal tasks in this project.
DFG Programme
Research Grants
Participating Persons
Dr. Gerald Dräger; Dr. Csaba Laszlo Sajti; Professor Dr.-Ing. Jochen Schein
