Coding transcriptome analyses reveal altered functions underlying immunotolerance of PEG-fused rat sciatic nerve allografts.
Coding transcriptome analyses reveal altered functions underlying immunotolerance of PEG-fused rat sciatic nerve allografts.
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编码转录组分析揭示了PEG融合大鼠坐骨神经同种异体移植物免疫耐受的潜在功能改变。
DOI：
10.1186/s12974-020-01953-8
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发表时间：
2020-10-02
影响因子：
9.3
通讯作者：
Bittner GD
中科院分区：
文献类型：
--
作者：
Smith TA;Ghergherehchi CL;Tucker HO;Bittner GD
目前使用周围神经同种异体移植物（PNAs）修复消融型周围神经损伤（PNI）的方法通常由于免疫排斥以及再生轴突芽生长缓慢和不准确而导致功能恢复不良。相比之下，消融型PNI修复的PNAs，使用一个多步骤的协议，其中一个步骤采用膜融合剂聚乙二醇（PEG），永久恢复坐骨神经介导的行为在几周内。PEG融合的PNA内的轴突和细胞保持活力，即使远交宿主和供体组织既不免疫抑制也不组织匹配。PEG融合的PNA表现出显着减少的T细胞和巨噬细胞浸润，主要组织相容性复合体I/II的表达和一贯的低凋亡。在这项研究中，我们分析了PEG融合PNA的编码转录组，以研究免疫抑制的可能机制。成年Sprague-Dawley大鼠的烧蚀型坐骨神经NIs使用PNAs和PEG融合方案结合神经缝合进行修复。电生理和行为测试证实了成功的PEG融合的PNA。RNA测序分析了PO后第14天PEG融合的PNA和阴性对照PNA（未用PEG处理）之间蛋白质编码基因的差异表达谱，沿着未操作的对照神经。测序结果通过定量逆转录PCR（RT-qPCR）进行验证，在某些情况下，免疫组织化学。PEG融合的PNA显示与先天性和适应性同种异体排斥反应相关的许多基因转录物的显着下调。雪旺细胞相关的转录物通常被上调，并且细胞过程如细胞外基质重塑和细胞/组织发育特别丰富。编码几种潜在免疫抑制蛋白的转录物（例如，血小板反应蛋白1和2）也在PEG融合的PNA中上调。这项研究是第一个表征编码转录组的PEG融合的PNA和确定细胞外基质的改变和抑制同种异体排斥反应之间的可能联系。这些结果为了解PEG介导的免疫抑制机制奠定了初步的分子基础。
Current methods to repair ablation-type peripheral nerve injuries (PNIs) using peripheral nerve allografts (PNAs) often result in poor functional recovery due to immunological rejection as well as to slow and inaccurate outgrowth of regenerating axonal sprouts. In contrast, ablation-type PNIs repaired by PNAs, using a multistep protocol in which one step employs the membrane fusogen polyethylene glycol (PEG), permanently restore sciatic-mediated behaviors within weeks. Axons and cells within PEG-fused PNAs remain viable, even though outbred host and donor tissues are neither immunosuppressed nor tissue matched. PEG-fused PNAs exhibit significantly reduced T cell and macrophage infiltration, expression of major histocompatibility complex I/II and consistently low apoptosis. In this study, we analyzed the coding transcriptome of PEG-fused PNAs to examine possible mechanisms underlying immunosuppression. Ablation-type sciatic PNIs in adult Sprague-Dawley rats were repaired using PNAs and a PEG-fusion protocol combined with neurorrhaphy. Electrophysiological and behavioral tests confirmed successful PEG-fusion of PNAs. RNA sequencing analyzed differential expression profiles of protein-coding genes between PEG-fused PNAs and negative control PNAs (not treated with PEG) at 14 days PO, along with unoperated control nerves. Sequencing results were validated by quantitative reverse transcription PCR (RT-qPCR), and in some cases, immunohistochemistry. PEG-fused PNAs display significant downregulation of many gene transcripts associated with innate and adaptive allorejection responses. Schwann cell-associated transcripts are often upregulated, and cellular processes such as extracellular matrix remodeling and cell/tissue development are particularly enriched. Transcripts encoding several potentially immunosuppressive proteins (e.g., thrombospondins 1 and 2) also are upregulated in PEG-fused PNAs. This study is the first to characterize the coding transcriptome of PEG-fused PNAs and to identify possible links between alterations of the extracellular matrix and suppression of the allorejection response. The results establish an initial molecular basis to understand mechanisms underlying PEG-mediated immunosuppression.
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影响因子：
4.2
作者：
Bittner GD;Sengelaub DR;Trevino RC;Peduzzi JD;Mikesh M;Ghergherehchi CL;Schallert T;Thayer WP
通讯作者：
Thayer WP
David Gene功能分类工具：一种以生物模块为中心的新型算法，可在功能上分析大基因列表。
影响因子：
12.3
作者：
Huang, Da Wei;Sherman, Brad T;Tan, Qina;Collins, Jack R;Alvord, W Gregory;Roayaei, Jean;Stephens, Robert;Baseler, Michael W;Lane, H Clifford;Lempicki, Richard A
通讯作者：
Lempicki, Richard A
影响因子：
3.3
作者：
Defrancesco-Lisowitz, A.;Lindborg, J. A.;Niemi, J. P.;Zigmond, R. E.
通讯作者：
Zigmond, R. E.
影响因子：
5
作者：
Chistiakov, Dimitry A.;Killingsworth, Murry C.;Bobryshev, Yuri V.
通讯作者：
Bobryshev, Yuri V.
影响因子：
6.2
作者：
Jiang, HS;Wynn, C;Kobayashi, M
通讯作者：
Kobayashi, M