Factor 3 regulates airway engraftment by human bronchial basal cells.

IF 5.4 2区 医学 Q1 CELL & TISSUE ENGINEERING
Susan D Reynolds, Cynthia L Hill, Alfahdah Alsudayri, Jacob T Stack, Kimberly M Shontz, Gianni Carraro, Barry R Stripp, Tendy Chiang
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Abstract

Cystic fibrosis transmembrane conductance regulator (CFTR) gene editing and transplantation of CFTR-gene corrected airway basal cells has the potential to cure CF lung disease. Although mouse studies established that cell transplantation was feasible, the engraftment rate was typically low and frequently less than the estimated therapeutic threshold. The purpose of this study was to identify genes and culture conditions that regulate the therapeutic potential of human bronchial basal cells. Factor 3 (F3, Tissue Factor 1) is a component of the extrinsic coagulation pathway and activates a cascade of proteases that convert fibrinogen to fibrin. Based on reports that F3 was necessary for human basal cell survival and adhesion in vitro, the present study evaluated F3 as a potential determinant of therapeutic fitness. The gene expression profile of F3 mRNA-positive human bronchial basal cells was evaluated by scRNAseq and the impact of the lung environment on F3 expression was modeled by varying in vitro culture conditions. F3 necessity for adhesion, proliferation, and differentiation was determined by CRISPR/Cas9 knockout (KO) of the F3 gene. Finally, the impact of F3 manipulation on engraftment was determined by orthotropic co-transplantation of wild-type and F3-KO cells into the airways of immunocompromised mice. In contrast with the hypothesis that F3 increases the therapeutic fitness of basal cells, F3 expression decreased engraftment. These studies guide the ongoing development of cellular therapies by showing that in vitro assessments may not predict therapeutic potential and that the lung milieu influences the functional properties of transplanted bronchial basal cells.

因子 3 调节人类支气管基底细胞的气道移植。
囊性纤维化跨膜传导调节器(CFTR)基因编辑和移植经 CFTR 基因校正的气道基底细胞有可能治愈 CF 肺病。虽然小鼠研究证实细胞移植是可行的,但移植率通常很低,经常低于估计的治疗阈值。本研究的目的是找出调节人类支气管基底细胞治疗潜力的基因和培养条件。因子 3(F3,组织因子 1)是外凝血途径的一个组成部分,能激活一连串蛋白酶,将纤维蛋白原转化为纤维蛋白。据报道,F3 是人类基底细胞在体外存活和粘附的必要条件,因此本研究将 F3 作为决定治疗效果的潜在因素进行评估。通过 scRNAseq 评估了 F3 mRNA 阳性的人支气管基底细胞的基因表达谱,并通过改变体外培养条件模拟了肺部环境对 F3 表达的影响。通过 CRISPR/Cas9 敲除(KO)F3 基因,确定了 F3 对粘附、增殖和分化的必要性。最后,通过将野生型和 F3-KO 细胞正交联合移植到免疫缺陷小鼠的气道中,确定了 F3 操作对移植的影响。与F3能提高基础细胞治疗能力的假设相反,F3的表达降低了移植效果。这些研究表明,体外评估可能无法预测治疗潜力,而肺部环境会影响移植支气管基底细胞的功能特性,从而为正在进行的细胞疗法开发提供指导。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Stem Cells Translational Medicine
Stem Cells Translational Medicine CELL & TISSUE ENGINEERING-
CiteScore
12.90
自引率
3.30%
发文量
140
审稿时长
6-12 weeks
期刊介绍: STEM CELLS Translational Medicine is a monthly, peer-reviewed, largely online, open access journal. STEM CELLS Translational Medicine works to advance the utilization of cells for clinical therapy. By bridging stem cell molecular and biological research and helping speed translations of emerging lab discoveries into clinical trials, STEM CELLS Translational Medicine will help move applications of these critical investigations closer to accepted best patient practices and ultimately improve outcomes. The journal encourages original research articles and concise reviews describing laboratory investigations of stem cells, including their characterization and manipulation, and the translation of their clinical aspects of from the bench to patient care. STEM CELLS Translational Medicine covers all aspects of translational cell studies, including bench research, first-in-human case studies, and relevant clinical trials.
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