Editing TINF2 as a potential therapeutic approach to restore telomere length in dyskeratosis congenita.
Editing TINF2 as a potential therapeutic approach to restore telomere length in dyskeratosis congenita.
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DOI：
10.1182/blood.2021013750
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发表时间：
2022-08-11
期刊：
影响因子：
20.3
通讯作者：
Hockemeyer, Dirk
中科院分区：
文献类型：
--
作者：
Choo, Seunga;Lorbeer, Franziska K.;Regalado, Samuel G.;Short, Sarah B.;Wu, Shannon;Rieser, Gabrielle;Bertuch, Alison A.;Hockemeyer, Dirk
在TINF2-DC等基因人类胚胎干细胞模型中，去除显性疾病等位基因可以恢复端粒长度。基因编辑的人类造血干细胞异种移植可用于疾病模型和测试潜在的治疗方法。编码保护素蛋白TIN2的TINF2基因突变会导致端粒缩短和遗传性骨髓(BM)衰竭综合征先天性角化不良(DC)。缺乏合适的模型系统限制了对干细胞中端粒缩短的机制的理解，从而阻碍了骨髓衰竭治疗选择的发展。在这里，我们在人胚胎干细胞(HESCs)和人造血干细胞(HSPC)中内源性引入TIN2-DC突变，以剖析疾病机制并确定拯救疾病表型的基因编辑策略。具有T284R疾病突变的hESCs表现出DC患者的短端粒表型。然而，突变的hESCs中的端粒在分化为端粒酶阴性细胞时并没有触发端粒的DNA损伤反应，也没有表现出加剧的端粒缩短。通过在外显子2中引入移码突变来破坏突变的TINF2等位基因，可以恢复干细胞的端粒长度和分化细胞的复制潜力。同样，我们在人类HSPC中引入了TIN2-DC疾病变异体来评估端粒长度和增殖能力的变化。最后，我们发现，在TINF2-DC患者HSPC中，可以在TINF2-DC患者HSPC中产生恢复hESCs端粒长度的TINF2外显子2的编辑。我们的研究证明了一种简单的基因干预方法，可以挽救干细胞中的TIN2-DC疾病表型，并提供了一个通用的平台来评估体内潜在治疗方法的有效性。
Removal of the dominant disease allele restores telomere length in an isogenic human embryonic stem cell model of TINF2-DC. Xenotransplantation of gene-edited human hematopoietic stem cells can be used to model disease and test potential therapeutic approaches. Mutations in the TINF2 gene, encoding the shelterin protein TIN2, cause telomere shortening and the inherited bone marrow (BM) failure syndrome dyskeratosis congenita (DC). A lack of suitable model systems limits the mechanistic understanding of telomere shortening in the stem cells and thus hinders the development of treatment options for BM failure. Here, we endogenously introduced TIN2-DC mutations in human embryonic stem cells (hESCs) and human hematopoietic stem and progenitor cells (HSPCs) to dissect the disease mechanism and identify a gene-editing strategy that rescued the disease phenotypes. The hESCs with the T284R disease mutation exhibited the short telomere phenotype observed in DC patients. Yet, telomeres in mutant hESCs did not trigger DNA damage responses at telomeres or show exacerbated telomere shortening when differentiated into telomerase-negative cells. Disruption of the mutant TINF2 allele by introducing a frameshift mutation in exon 2 restored telomere length in stem cells and the replicative potential of differentiated cells. Similarly, we introduced TIN2-DC disease variants in human HSPCs to assess the changes in telomere length and proliferative capacity. Lastly, we showed that editing at exon 2 of TINF2 that restored telomere length in hESCs could be generated in TINF2-DC patient HSPCs. Our study demonstrates a simple genetic intervention that rescues the TIN2-DC disease phenotype in stem cells and provides a versatile platform to assess the efficacy of potential therapeutic approaches in vivo.
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影响因子：
16.6
作者：
Lai TP;Zhang N;Noh J;Mender I;Tedone E;Huang E;Wright WE;Danuser G;Shay JW
通讯作者：
Shay JW
影响因子：
64.8
作者：
Batista, Luis F. Z.;Pech, MatthewF.;Zhong, Franklin L.;Nguyen, Ha Nam;Xie, Kathleen T.;Zaug, Arthur J.;Crary, Sharon M.;Choi, Jinkuk;Sebastiano, Vittorio;Cherry, Athena;Giri, Neelam;Wernig, Marius;Alter, Blanche P.;Cech, Thomas R.;Savage, Sharon A.;Pera, Renee A. Reijo;Artandi, Steven E.
通讯作者：
Artandi, Steven E.
DOI：
10.1182/asheducation-2011.1.480
发表时间：
2011-12-01
期刊：
HEMATOLOGY-AMERICAN SOCIETY HEMATOLOGY EDUCATION PROGRAM
影响因子：
--
作者：
Dokal, Inderjeet
通讯作者：
Dokal, Inderjeet
影响因子：
6.6
作者：
He, Huiling;Li, Wei;de la Chapelle, Albert
通讯作者：
de la Chapelle, Albert
影响因子：
30.8
作者：
Baird, DM;Rowson, J;Kipling, D
通讯作者：
Kipling, D