A 2020 study identified AAV2 and AAV serotype 6 (AAV6) as having the highly efficient transduction of the human lung parenchyma

A 2020 study identified AAV2 and AAV serotype 6 (AAV6) as having the highly efficient transduction of the human lung parenchyma. corrective strategies. transfer of a functional copy of has been envisioned as a CF airway treatment since 1989 when the gene was identified as the cause of this multisystemic disease (Tsui et al., 1985; Wainwright et al., 1985). Gene therapy has received FDA approval for treatment of monogenic disorders (U.S. Food and Drug Administration. 2020) such as spinal muscular atrophy (Kariyawasam et al., 2018), coagulative disorders (Batty and Lillicrap 2019), and immunodeficiency diseases DMCM hydrochloride (Booth et al., 2019), but not yet for CF. Numerous research programs and clinical trials have been undertaken to explicate the most effective vector (viral or non-viral) to deliver to airway cells (Griesenbach et al., 2015). However, clinical efficacy of these vectors in humans has been insignificant and inconsistent in improving lung function (Alton et al., 2015a). The greatest barrier to enabling clinical translation of gene therapy for CF remains the lack of an effective delivery system to the lungs. A successful gene therapy system for restoration of CFTR function needs to navigate the complexities of the lung clearance and innate immunity defense functions that are further complicated in the CF airways due to increased mucus volume and viscosity (reviewed in (Donnelley and Parsons 2018)). Even if these obstacles are circumvented, heterogeneous and highly regulated CFTR expression in various cell types of the lung raises the question of the most appropriate cellular target. One proposed strategy to deal with the challenges associated with delivery of to the airway cells is to correct the airway cells followed by transplanting the corrected cells to repopulate the patients lung with hematopoietic stem cell gene therapy, Strimvelis, which was approved for treatment of adenosine deaminase-severe combined immunodeficiency (Stirnadel-Farrant et al., 2018). In this review, we will first describe alternative strategies to CFTR DNA therapy, and discuss the advances in the main groups of viral and non-viral vectors that have shown promise in CF therapy. The second part of this review will focus on DMCM hydrochloride recent progress in cell-based therapies, including the gene editing technologies that facilitate CFTR correction in cellsin the collected cells by a) addition or b) editing strategies. EGF 3) The CFTR-corrected regenerative cells are expanded to reach a therapeutic dose, and then 4) transplanted back to repopulate the patient lung epithelium. Therapeutic Genetic Material Other Than DNA: RNA Addition and Repair The earliest efforts to deliver genetic material into diseased cells focused on directly introducing therapeutic DNA as an addition strategy to subsequently produce functional CFTR protein (reviewed in (Cooney et al., 2018)). A novel alternative to DNA therapeutics is based on addition of RNA. Since the functional site of messenger RNA (mRNA) is the cell cytoplasm, the challenge of nuclear translocation is eliminated (Hajj and Whitehead 2017). Exogenous nucleic acids are susceptible to degradation by nucleases and can trigger DMCM hydrochloride an immune response upon cellular entry (Alexopoulou et al., 2001; Kariko et al., 2004). Therefore, current strategies utilize chemical modification of the nucleic acid bases to reduce immunogenicity and increase stability (Sahin et al., 2014; Pardi et al., 2015). Manufacturing and addition of modifications to RNA is easier than DNA, extending the usefulness of RNA therapy (Kuhn et al., 2012). Yet, repeat DMCM hydrochloride RNA administration remains necessary to sustain therapeutic levels of protein (Patel et al., 2019b). Successful delivery of chemically modified CFTR mRNA to patient-derived bronchial epithelial cells has demonstrated DMCM hydrochloride increased CFTR expression at the plasma membrane and rescue of.