Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, University of Southern California Keck School of Medicine, Los Angeles, CA 90033, USA.
Institute for Systems Genetics, NYU Langone Health, New York, NY 10016, USA; Department of Biochemistry and Molecular Pharmacology, NYU Grossman School of Medicine, New York, NY 10016, USA.
Med. 2024 Aug 9;5(8):1016-1029.e4. doi: 10.1016/j.medj.2024.05.003. Epub 2024 May 21.
Xenotransplantation of genetically engineered porcine organs has the potential to address the challenge of organ donor shortage. Two cases of porcine-to-human kidney xenotransplantation were performed, yet the physiological effects on the xenografts and the recipients' immune responses remain largely uncharacterized.
We performed single-cell RNA sequencing (scRNA-seq) and longitudinal RNA-seq analyses of the porcine kidneys to dissect xenotransplantation-associated cellular dynamics and xenograft-recipient interactions. We additionally performed longitudinal scRNA-seq of the peripheral blood mononuclear cells (PBMCs) to detect recipient immune responses across time.
Although no hyperacute rejection signals were detected, scRNA-seq analyses of the xenografts found evidence of endothelial cell and immune response activation, indicating early signs of antibody-mediated rejection. Tracing the cells' species origin, we found human immune cell infiltration in both xenografts. Human transcripts in the longitudinal bulk RNA-seq revealed that human immune cell infiltration and the activation of interferon-gamma-induced chemokine expression occurred by 12 and 48 h post-xenotransplantation, respectively. Concordantly, longitudinal scRNA-seq of PBMCs also revealed two phases of the recipients' immune responses at 12 and 48-53 h. Lastly, we observed global expression signatures of xenotransplantation-associated kidney tissue damage in the xenografts. Surprisingly, we detected a rapid increase of proliferative cells in both xenografts, indicating the activation of the porcine tissue repair program.
Longitudinal and single-cell transcriptomic analyses of porcine kidneys and the recipient's PBMCs revealed time-resolved cellular dynamics of xenograft-recipient interactions during xenotransplantation. These cues can be leveraged for designing gene edits and immunosuppression regimens to optimize xenotransplantation outcomes.
This work was supported by NIH RM1HG009491 and DP5OD033430.
基因工程猪器官的异种移植具有解决器官供体短缺挑战的潜力。已经进行了两例猪到人体肾脏异种移植，但异种移植物和受者免疫反应的生理影响在很大程度上仍未得到充分描述。
我们对供体猪的肾脏进行了单细胞 RNA 测序（scRNA-seq）和纵向 RNA-seq 分析，以剖析异种移植相关的细胞动力学和移植物-受者相互作用。我们还对外周血单核细胞（PBMC）进行了纵向 scRNA-seq 分析，以检测随时间推移的受者免疫反应。
尽管没有检测到超急性排斥反应信号，但异种移植物的 scRNA-seq 分析发现了内皮细胞和免疫反应激活的证据，表明存在抗体介导的排斥反应的早期迹象。追踪细胞的种属来源，我们发现异种移植物中存在人类免疫细胞浸润。纵向 bulk RNA-seq 中的人类转录本表明，人类免疫细胞浸润和干扰素-γ诱导的趋化因子表达的激活分别在异种移植后 12 和 48 小时发生。相应地，PBMC 的纵向 scRNA-seq 也揭示了受者免疫反应的两个阶段，分别在 12 和 48-53 小时。最后，我们在异种移植物中观察到与异种移植相关的肾脏组织损伤的全基因组表达特征。令人惊讶的是，我们在两个异种移植物中都检测到增殖细胞的快速增加，这表明猪组织修复程序的激活。
对供体猪的肾脏和受者的 PBMC 进行纵向和单细胞转录组分析揭示了异种移植过程中移植物-受者相互作用的时间分辨细胞动力学。这些线索可用于设计基因编辑和免疫抑制方案，以优化异种移植的结果。
这项工作得到了 NIH RM1HG009491 和 DP5OD033430 的支持。
