Low immunogenic endothelial cells endothelialize the Left Ventricular Assist Device
Low immunogenic endothelial cells endothelialize the Left Ventricular Assist Device
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DOI：
10.1038/s41598-019-47780-7
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发表时间：
2019-08
影响因子：
4.6
通讯作者：
C. Figueiredo;Dorothee Eicke;Yuliia Yuzefovych;M. Avşar;J. Hanke;M. Pflaum;J. Schmitto;R. Blasczyk;A. Haverich;B. Wiegmann
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作者：
C. Figueiredo;Dorothee Eicke;Yuliia Yuzefovych;M. Avşar;J. Hanke;M. Pflaum;J. Schmitto;R. Blasczyk;A. Haverich;B. Wiegmann
左心室辅助装置（LVAD）表面的血液相容性低，需要进行抗凝治疗。内皮细胞（EC）接种可以支持血液相容性，然而，自体EC的可用性是有限的。相比之下，同种异体EC容易获得足够的量，但HLA差异诱导有害的免疫应答，导致EC损失。在这项研究中，我们研究了使用同种异体低免疫原性EC内皮化LVAD烧结流入套管（SIC）的可行性。为了降低内皮细胞的免疫原性，我们应用诱导型慢病毒载体来递送短发夹RNA以沉默HLA I类表达。内皮细胞上的HLA I类表达被有条件地沉默高达70%。用HLA表达或HLA沉默的EC实现了足够的和相当的内皮化速率。细胞与烧结流入插管（SIC）接触或沉默HLA表达不会损害细胞增殖。内皮细胞表型和血栓形成标志物或细胞因子分泌谱的水平保持不受影响。HLA沉默的EC涂覆的SIC表现出降低的促凝性。与天然EC相比，HLA沉默的EC在暴露于同种异体T细胞或特异性抗HLA抗体时显示出较低的细胞裂解率。同种异体HLA沉默的EC可能成为LVAD内皮化的有价值的来源，以降低免疫原性，并相应地降低可能引起严重副作用的抗凝治疗的需要。
Low haemocompatibility of left ventricular assist devices (LVAD) surfaces necessitates anticoagulative therapy. Endothelial cell (EC) seeding can support haemocompatibility, however, the availability of autologous ECs is limited. In contrast, allogeneic ECs are readily available in sufficient quantity, but HLA disparities induce harmful immune responses causing EC loss. In this study, we investigated the feasibility of using allogeneic low immunogenic ECs to endothelialize LVAD sintered inflow cannulas (SIC). To reduce the immunogenicity of ECs, we applied an inducible lentiviral vector to deliver short-hairpins RNA to silence HLA class I expression. HLA class I expression on ECs was conditionally silenced by up to 70%. Sufficient and comparable endothelialization rates were achieved with HLA-expressing or HLA-silenced ECs. Cell proliferation was not impaired by cell-to-Sintered Inflow Cannulas (SIC) contact or by silencing HLA expression. The levels of endothelial phenotypic and thrombogenic markers or cytokine secretion profiles remained unaffected. HLA-silenced ECs-coated SIC exhibited reduced thrombogenicity. In contrast to native ECs, HLA-silenced ECs showed lower cell lysis rates when exposed to allogeneic T cells or specific anti-HLA antibodies. Allogeneic HLA-silenced ECs could potentially become a valuable source for LVAD endothelialization to reduce immunogenicity and correspondingly the need for anticoagulative therapy which can entail severe side effects.