title: In Vitro Evaluation of a Novel mRNA-Based Therapeutic Strategy for the Treatment of Patients Suffering from Alpha-1-Antitrypsin Deficiency.
abstract: In single-gene disorders, like alpha-1-antitrypsin deficiency (AATD), a gene mutation causes missing or dysfunctional protein synthesis. This, in turn, can lead to serious complications for the patient affected. Furthermore, single-gene disorders are associated with severe early-onset conditions and necessitate expensive lifelong care. Until nowadays, therapeutic treatment options are still limited, cost-intensive, or lack effectiveness. For these reasons, we aim to develop a novel mRNA-based therapeutic strategy for the treatment of single-gene disorders, such as AATD, which is based on the induction of de novo synthesis of the functional proteins. Therefore, an alpha-1-antitrypsin (AAT) encoding mRNA was generated by in vitro transcription. After in vitro delivery of the mRNA to different cells, protein expression and functionality, as well as adverse effects and mRNA serum stability, were analyzed. Our results show that the AAT mRNA-transfected cells express the AAT protein in high amounts within the first 24 h. Moreover, the expressed AAT protein is highly functional, since the activity of elastase is significantly inhibited. Our data also show that mRNA concentrations up to 1 μg per 150,000 cells have no adverse effects on cell viability and immune activation. Furthermore, the encapsulated AAT encoding mRNA is stable and functional in human serum for up to 30 min. Overall, the proposed project provides an innovative, highly promising, and safe therapeutic approach and, thus, promises a novel progress in the treatment of single-gene disorders, whereby affected patients could greatly benefit.
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Tatjana
authors.initials: T
authors.lastname: Michel
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Anna
authors.initials: A
authors.lastname: Kankura
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Martha L
authors.initials: ML
authors.lastname: Salinas Medina
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Julia
authors.initials: J
authors.lastname: Kurz
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Andreas
authors.initials: A
authors.lastname: Behring
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Meltem
authors.initials: M
authors.lastname: Avci-Adali
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Andrea
authors.initials: A
authors.lastname: Nolte
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Christian
authors.initials: C
authors.lastname: Schlensak
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Hans Peter
authors.initials: HP
authors.lastname: Wendel
authors.affiliation: Clinical Research Laboratory, Department of Thoracic and Cardiovascular Surgery, University Hospital Tuebingen, Tuebingen University , Tuebingen, Germany .
authors.forename: Stefanie
authors.initials: S
authors.lastname: Krajewski
mesh_terms: D016207:Cytokines / Q000096:biosynthesis; D015316:Genetic Therapy; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D014176:Protein Biosynthesis; D020871:RNA Stability; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D019896:alpha 1-Antitrypsin Deficiency / Q000628:therapy*
chemical_list: D016207:Cytokines; D012333:RNA, Messenger