Phenotypic Characterization of Transgenic Mice Expressing Human IGFBP-5.
Phenotypic Characterization of Transgenic Mice Expressing Human IGFBP-5.
复制标题
DOI：
10.3390/ijms22010335
复制
发表时间：
2020-12-30
影响因子：
5.6
通讯作者：
Feghali-Bostwick C
中科院分区：
文献类型：
--
作者：
Nguyen XX;Sanderson M;Helke K;Feghali-Bostwick C
肺纤维化是系统性硬化症（SSc）和特发性肺纤维化（IPF）等纤维增殖性疾病发病和死亡的重要原因之一。胰岛素样生长因子结合蛋白 5 (IGFBP-5) 是 IGFBP 蛋白家族的保守成员，在 SSc 和 IPF 肺组织中过度表达。在这项研究中，我们使用转基因模型研究了 IGFBP-5 在体内纤维化发展中的功能作用。我们使用 CRISPR/Cas9 敲入技术培育出普遍表达人类 IGFBP-5 的转基因小鼠。我们的数据表明，杂合子和纯合子小鼠均能存活并表达人 IGFBP-5 (hIGFBP-5)。转基因小鼠的细胞外基质 (ECM) 基因表达增加，尤其是表达较高转基因水平的小鼠的肺和皮肤组织中的 Col3a1、Fn 和 Lox。皮肤组织的组织学分析显示，与野生型小鼠相比，杂合子和纯合子小鼠的真皮厚度增加，肺部组织学显示出细微的变化。这些变化在表达较高水平 hIGFBP-5 的动物中更为明显。博来霉素增加了野生型小鼠中的 ECM 基因表达，并加剧了转基因小鼠中 ECM 基因表达的增加，表明转基因表达加剧了博来霉素诱导的肺纤维化。从纯合转基因小鼠肺组织培养的原代肺成纤维细胞显示出 ECM 基因表达和蛋白质水平显着增加，进一步支持了 IGFBP-5 导致成纤维细胞纤维化表型的观察结果。总之，表达人IGFBP-5的转基因小鼠可以作为检查IGFBP-5体内功能的有用动物模型。
Pulmonary fibrosis is one of the important causes of morbidity and mortality in fibroproliferative disorders such as systemic sclerosis (SSc) and idiopathic pulmonary fibrosis (IPF). Insulin-like growth factor binding protein-5 (IGFBP-5) is a conserved member of the IGFBP family of proteins that is overexpressed in SSc and IPF lung tissues. In this study, we investigated the functional role of IGFBP-5 in the development of fibrosis in vivo using a transgenic model. We generated transgenic mice ubiquitously expressing human IGFBP-5 using CRISPR/Cas9 knock-in. Our data show that the heterozygous and homozygous mice are viable and express human IGFBP-5 (hIGFBP-5). Transgenic mice had increased expression of extracellular matrix (ECM) genes, especially Col3a1, Fn, and Lox in lung and skin tissues of mice expressing higher transgene levels. Histologic analysis of the skin tissues showed increased dermal thickness, and the lung histology showed subtle changes in the heterozygous and homozygous mice as compared with the wild-type mice. These changes were more pronounced in animals expressing higher levels of hIGFBP-5. Bleomycin increased ECM gene expression in wild-type mice and accentuated an increase in ECM gene expression in transgenic mice, suggesting that transgene expression exacerbated bleomycin-induced pulmonary fibrosis. Primary lung fibroblasts cultured from lung tissues of homozygous transgenic mice showed significant increases in ECM gene expression and protein levels, further supporting the observation that IGFBP-5 resulted in a fibrotic phenotype in fibroblasts. In summary, transgenic mice expressing human IGFBP-5 could serve as a useful animal model for examining the function of IGFBP-5 in vivo.
登录
查看更多内容
Htnfalpha转基因小鼠的广泛表型表征。
影响因子：
--
作者：
Hayward, Michael D;Jones, Beverly K;Saparov, Arman;Hain, Heather S;Trillat, Anne-Cecile;Bunzel, Michelle M;Corona, Aaron;Li-Wang, Bifang;Strenkowski, Bryan;Giordano, Caroline;Shen, Hai;Arcamone, Emily;Weidlick, Jeffrey;Vilensky, Maria;Tugusheva, Marina;Felkner, Roland H;Campbell, William;Rao, Yu;Grass, David S;Buiakova, Olesia
通讯作者：
Buiakova, Olesia
影响因子：
3.6
作者：
Kotarkonda, Lakshmi Kanth;Kulshrestha, Ritu;Ravi, Krishnan
通讯作者：
Ravi, Krishnan
影响因子：
2.9
作者：
Mohan, S;Thompson, GR;Baylink, DJ
通讯作者：
Baylink, DJ
影响因子：
--
作者：
Hsu, Eileen;Shi, Haiwen;Jordan, Rick M.;Lyons-Weiler, James;Pilewski, Joseph M.;Feghali-Bostwick, Carol A.
通讯作者：
Feghali-Bostwick, Carol A.
影响因子：
6
作者：
Pilewski, JM;Liu, LX;Feghali-Bostwick, CA
通讯作者：
Feghali-Bostwick, CA