Mao Mao, Ishikawa Yoshihiro, Labelle-Dumais Cassandre, Wang Xiaowei, Kuo Yien-Ming, Gaffney Uma B, Smith Megan E, Abdala Carlie N, Lebedev Matthew D, Paradee William J, Gould Douglas B
Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.
Genome Editing Core Facility, University of Iowa, Iowa City, IA, USA.
J Cell Biol. 2025 Jun 2;224(6). doi: 10.1083/jcb.202409153. Epub 2025 Apr 25.
Basement membranes (BMs) are specialized extracellular matrix (ECM) structures essential for organ morphogenesis, architecture, and function. BM composition and properties vary between tissues, developmental stages, and disease states, and there is only a rudimentary understanding of BM dynamics. Here, we introduce a versatile mouse model carrying a multifunctional dual-color fluorescence tagged allele with knockout potential for the fundamental BM component type IV collagen alpha 1 (COL4A1). This allele enables the characterization of cell type- and time-specific contributions to BMs and the generation of a conditional Col4a1 null allele. We demonstrate the utility of this unique genetic resource in providing clinically relevant insights for individuals with Gould syndrome - a multisystem disorder caused by COL4A1 and COL4A2 mutations. We show active COL4A1 turnover in postnatal cerebrovascular BMs, identifying a potential interventional window for cerebrovascular manifestations associated with Gould syndrome. We also demonstrate that heterozygous Col4a1 deletion is significantly less pathogenic than dominant Col4a1 missense mutations, which has important implications for gene therapy.
基底膜（BMs）是特殊的细胞外基质（ECM）结构，对器官形态发生、结构和功能至关重要。BM的组成和特性在不同组织、发育阶段和疾病状态之间存在差异，人们对BM动态变化的了解还很初步。在此，我们介绍一种多功能小鼠模型，它携带一个多功能双色荧光标记等位基因，对基本的BM成分IV型胶原蛋白α1（COL4A1）具有敲除潜力。该等位基因能够表征细胞类型和时间特异性对BMs的贡献，并产生条件性Col4a1无效等位基因。我们展示了这种独特遗传资源的效用，为患有古尔德综合征（一种由COL4A1和COL4A2突变引起的多系统疾病）的个体提供临床相关见解。我们显示出生后脑血管BMs中有活跃的COL4A1更新，确定了与古尔德综合征相关的脑血管表现的潜在干预窗口。我们还证明，杂合性Col4a1缺失的致病性明显低于显性Col4a1错义突变，这对基因治疗具有重要意义。
