King Abdullah International Medical Research Center (KAIMRC), King Saud Bin Abdulaziz University for Health Sciences, Ministry of National Guard for Health Affairs, Jeddah, Saudi Arabia.
Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal, Saudi Arabia.
Stem Cell Res Ther. 2023 Dec 18;14(1):374. doi: 10.1186/s13287-023-03612-0.
Human iPSCs' derivation and use in clinical studies are transforming medicine. Yet, there is a high cost and long waiting time associated with autologous iPS-based cellular therapy, and the genetic engineering of hypo-immunogenic iPS cell lines is hampered with numerous hurdles. Therefore, it is increasingly interesting to create cell stocks based on HLA haplotype distribution in a given population. This study aimed to assess the potential of HLA-based iPS banking for the Saudi population.
In this study, we interrogated the HLA database of the Saudi Stem Cell Donor Registry (SSCDR), containing high-resolution HLA genotype data from 64,315 registered Saudi donors at the time of analysis. This database was considered to be a representative sample of the Saudi population. The most frequent HLA haplotypes in the Saudi population were determined, and an in-house developed iterative algorithm was used to identify their HLA matching percentages in the SSCDR database and cumulative coverage. Subsequently, to develop a clinically relevant protocol for iPSCs generation, and to illustrate the applicability of the concept of HLA-based banking for cell therapy purposes, the first HLA-based iPS cell line in Saudi Arabia was generated. Clinically relevant methods were employed to generate the two iPS clones from a homozygous donor for the most prevalent HLA haplotype in the Saudi population. The generated lines were then assessed for pluripotency markers, and their ability to differentiate into all three germ layers, beating cardiomyocytes, and neural progenitors was examined. Additionally, the genetic stability of the HLA-iPS cell lines was verified by comparing the mutational burden in the clones and the original blood sample, using whole-genome sequencing. The standards set by the American College of Medical Genetics and Genomics (ACMG) were used to determine the clinical significance of identified variants.
The analysis revealed that the establishment of only 13 iPSC lines would match 30% of the Saudi population, 39 lines would attain 50% coverage, and 596 lines would be necessary for over 90% coverage. The proof-of-concept HLA-iPSCs, which cover 6.1% of the Saudi population, successfully demonstrated pluripotency and the ability to differentiate into various cell types including beating cardiomyocytes and neuronal progenitors. The comprehensive genetic analysis corroborated that all identified variants in the derived iPSCs were inherently present in the original donor sample and were classified as benign according to the standards set by the ACMG.
Our study sets a road map for introducing iPS-based cell therapy in the Kingdom of Saudi Arabia. It underscores the pragmatic approach of HLA-based iPSC banking which circumvents the limitations of autologous iPS-based cellular therapies. The successful generation and validation of iPSC lines based on the most prevalent HLA haplotype in the Saudi population signify a promising step toward broadening the accessibility and applicability of stem cell therapies and regenerative medicine in Saudi Arabia.
人类诱导多能干细胞（iPSCs）的衍生和在临床研究中的应用正在改变医学。然而，自体 iPS 为基础的细胞治疗费用高昂且等待时间长，并且低免疫原性 iPS 细胞系的基因工程存在诸多障碍。因此，基于特定人群中 HLA 单倍型分布来创建细胞库越来越受到关注。本研究旨在评估基于 HLA 的 iPS 存储库在沙特人群中的应用潜力。
在这项研究中，我们分析了沙特干细胞捐献者登记处（SSCDR）的 HLA 数据库，该数据库包含了 64315 名已注册的沙特供体在分析时的高分辨率 HLA 基因型数据。该数据库被认为是沙特人群的代表性样本。确定了沙特人群中最常见的 HLA 单倍型，并使用内部开发的迭代算法来确定其在 SSCDR 数据库中的 HLA 匹配百分比和累积覆盖率。随后，为了开发临床相关的 iPSCs 生成方案，并说明基于 HLA 的存储库在细胞治疗目的方面的适用性，我们在沙特阿拉伯生成了第一个基于 HLA 的 iPS 细胞系。使用临床相关方法从最常见的 HLA 单倍型在沙特人群中的纯合供体中生成了两个 iPS 克隆。然后评估生成的细胞系的多能性标志物，并检查其分化为所有三个胚层、跳动的心肌细胞和神经祖细胞的能力。此外，通过比较克隆和原始血液样本的突变负担，使用全基因组测序来验证 HLA-iPSC 细胞系的遗传稳定性。使用美国医学遗传学与基因组学学院（ACMG）设定的标准来确定鉴定变体的临床意义。
分析表明，仅建立 13 个 iPSC 系就可以匹配 30%的沙特人口，39 个系可以达到 50%的覆盖率，而 596 个系则需要超过 90%的覆盖率。涵盖沙特人口 6.1%的概念验证 HLA-iPSCs 成功证明了其多能性和分化为各种细胞类型的能力，包括跳动的心肌细胞和神经元祖细胞。全面的遗传分析证实，衍生的 iPSCs 中所有鉴定的变体在原始供体样本中固有存在，并且根据 ACMG 设定的标准分类为良性。
我们的研究为在沙特阿拉伯引入基于 iPS 的细胞治疗制定了路线图。它强调了基于 HLA 的 iPSC 存储库的务实方法，该方法规避了自体 iPS 为基础的细胞治疗的局限性。基于沙特人群中最常见的 HLA 单倍型成功生成和验证 iPSC 系，标志着朝着扩大干细胞治疗和再生医学在沙特阿拉伯的可及性和适用性迈出了有希望的一步。
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