Targeted Gene Modification of Hematopoietic Progenitor Cells in Mice Following Systemic Administration of a PNA-peptide Conjugate
Targeted Gene Modification of Hematopoietic Progenitor Cells in Mice Following Systemic Administration of a PNA-peptide Conjugate
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DOI：
10.1038/mt.2011.163
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发表时间：
2012-01-01
影响因子：
12.4
通讯作者：
Glazer, Peter M.
中科院分区：
文献类型：
--
作者：
Rogers, Faye A.;Lin, Sharon S.;Glazer, Peter M.
造血干细胞（HSC）基因治疗为各种血液病的新治疗方法的发展提供了希望。然而，HSC的有效体内修饰已被证明是具有挑战性的，因此对该方法的治疗潜力施加了限制。在此，我们提供了一种基因靶向策略，允许在小鼠的HSC中进行位点特异性的体内基因修饰。通过将三链体形成肽核酸（PNA）与转运肽Antennapedia（Antp）缀合，我们成功地实现了造血祖细胞的体内染色体基因组修饰，同时仍保留完整的分化能力。PNA-Antp偶联物全身给药后，在多个体细胞组织以及造血系统的多个区室中观察到序列特异性基因修饰，包括红细胞系、髓细胞系和淋巴细胞系。作为长期可再生HSC中基因靶向的真实功能测量，我们还证明了在移植来自PNA-Antp处理的供体小鼠的骨髓后，原代受体小鼠的骨髓和脾脏中保留的基因组修饰。我们的方法提供了一种微创替代离体基因治疗，通过消除对干细胞动员和收获，离体操作，干细胞移植的复杂步骤的需要。因此，我们的方法可能为单基因血液病（如镰状细胞性贫血和地中海贫血）的个体化治疗提供新的选择。
Hematopoietic stem cell (HSC) gene therapy offers promise for the development of new treatments for a variety of hematologic disorders. However, efficient in vivo modification of HSCs has proved challenging, thus imposing constraints on the therapeutic potential of this approach. Herein, we provide a gene-targeting strategy that allows site-specific in vivo gene modification in the HSCs of mice. Through conjugation of a triplex-forming peptide nucleic acid (PNA) to the transport peptide, antennapedia (Antp), we achieved successful in vivo chromosomal genomic modification of hematopoietic progenitor cells, while still retaining intact differentiation capabilities. Following systemic administration of PNA-Antp conjugates, sequence-specific gene modification was observed in multiple somatic tissues as well as within multiple compartments of the hematopoietic system, including erythroid, myeloid, and lymphoid cell lineages. As a true functional measure of gene targeting in a long-term renewable HSC, we also demonstrate preserved genomic modification in the bone marrow and spleen of primary recipient mice following transplantation of bone marrow from PNA-Antp-treated donor mice. Our approach offers a minimally invasive alternative to ex vivo gene therapy, by eliminating the need for the complex steps of stem cell mobilization and harvesting, ex vivo manipulation, and transplantation of stem cells. Therefore, our approach may provide new options for individualized therapies in the treatment of monogenic hematologic diseases such as sickle cell anemia and thalassemia.