用 CRISPR 碱基编辑方法对人类健康和衰老过程中端粒生物学紊乱相关基因进行功能评估

IF 4.4 4区 医学 Q1 GERIATRICS & GERONTOLOGY
Gustavo Borges, Yahya Benslimane, Lea Harrington
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引用次数: 0

摘要

端粒生物学疾病(TBDs)是一组以端粒过短和/或端粒功能障碍为特征的罕见疾病。它们包括一组骨髓衰竭综合征、特发性肺纤维化和肝病等疾病。负责端粒平衡的基因中的遗传变异(变体)与TBDs有关。尽管有许多变体已被确定为致病基因,但仍有更多的变体需要更好地了解。对 TBDs 的研究具有挑战性,因为这些变异因其罕见性而难以确定,很难预测它们对疾病发病的影响,而且也没有足够的样本可供研究。我们对致病变异的了解大多来自于对TBD患者及其亲属端粒酶活性的评估。然而,我们仍然缺乏一个基于细胞的模型来识别新的变异体,并研究这些变异体对TBDs相关基因的长期影响。在这里,我们提出了一种基于细胞的模型,利用 CRISPR 碱基编辑技术诱变参与端粒生物学的 21 个基因的内源等位基因。我们确定了编码 17 种不同蛋白质的基因中影响细胞生长的关键残基。我们为 TBDs 患者中意义不明的变异提供了功能性证据。我们还发现了对端粒酶抑制具有抗性的变体,这些变体与表达野生型端粒酶的细胞类似,在体外肿瘤生长试验中表现出更高的致瘤潜力。我们相信,这种基于细胞的方法将极大地推动我们对TBDs生物学的了解,并可能有助于开发治疗这类疾病的新疗法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A CRISPR base editing approach for the functional assessment of telomere biology disorder-related genes in human health and aging

A CRISPR base editing approach for the functional assessment of telomere biology disorder-related genes in human health and aging

Telomere Biology Disorders (TBDs) are a group of rare diseases characterized by the presence of short and/or dysfunctional telomeres. They comprise a group of bone marrow failure syndromes, idiopathic pulmonary fibrosis, and liver disease, among other diseases. Genetic alterations (variants) in the genes responsible for telomere homeostasis have been linked to TBDs. Despite the number of variants already identified as pathogenic, an even more significant number must be better understood. The study of TBDs is challenging since identifying these variants is difficult due to their rareness, it is hard to predict their impact on the disease onset, and there are not enough samples to study. Most of our knowledge about pathogenic variants comes from assessing telomerase activity from patients and their relatives affected by a TBD. However, we still lack a cell-based model to identify new variants and to study the long-term impact of such variants on the genes involved in TBDs. Herein, we present a cell-based model using CRISPR base editing to mutagenize the endogenous alleles of 21 genes involved in telomere biology. We identified key residues in the genes encoding 17 different proteins impacting cell growth. We provide functional evidence for variants of uncertain significance in patients with TBDs. We also identified variants resistant to telomerase inhibition that, similar to cells expressing wild-type telomerase, exhibited increased tumorigenic potential using an in vitro tumour growth assay. We believe that such cell-based approaches will significantly advance our understanding of the biology of TBDs and may contribute to the development of new therapies for this group of diseases.

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来源期刊
Biogerontology
Biogerontology 医学-老年医学
CiteScore
8.00
自引率
4.40%
发文量
54
审稿时长
>12 weeks
期刊介绍: The journal Biogerontology offers a platform for research which aims primarily at achieving healthy old age accompanied by improved longevity. The focus is on efforts to understand, prevent, cure or minimize age-related impairments. Biogerontology provides a peer-reviewed forum for publishing original research data, new ideas and discussions on modulating the aging process by physical, chemical and biological means, including transgenic and knockout organisms; cell culture systems to develop new approaches and health care products for maintaining or recovering the lost biochemical functions; immunology, autoimmunity and infection in aging; vertebrates, invertebrates, micro-organisms and plants for experimental studies on genetic determinants of aging and longevity; biodemography and theoretical models linking aging and survival kinetics.
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