Basova Liana V, Tang Xin, Umasume Takeshi, Gromova Anastasia, Zyrianova Tatiana, Shmushkovich Taisia, Wolfson Alexey, Hawley Dillon, Zoukhri Driss, Shestopalov Valery I, Makarenkova Helen P
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, United States.
Advirna LLC, Cambridge, Massachusetts, United States.
Invest Ophthalmol Vis Sci. 2017 Nov 1;58(13):5654-5665. doi: 10.1167/iovs.17-22071.
Sjögren's syndrome is a systemic chronic autoimmune inflammatory disease that primarily targets the salivary and lacrimal glands (LGs). Currently there is no cure; therefore, cell-based regenerative therapy may be a viable option. LG inflammation is facilitated by extracellular ATP and mediated by the Pannexin-1 (Panx1) membrane channel glycoprotein. We propose that suppression of inflammation through manipulation of Panx1 activity can stimulate epithelial cell progenitor (EPCP) engraftment.
The expression of pannexins in the mouse and human LG was assayed by qRT-PCR and immunostaining. Acute LG inflammation was induced by interleukin-1α (IL1α) injection. Prior to EPCP transplantation, IL1α-injured or chronically inflamed LGs of thrombospondin-1-null mice (TSP-1-/-) were treated with the Panx1-specific blocking peptide (10panx) or the self-deliverable RNAi (sdRNAi). The efficacy of cell engraftment and the area of inflammation were analyzed by microscopy.
Panx1 and Panx2 were detected in the mouse and human LGs. Panx1 and proinflammatory factors were upregulated during acute inflammation at days 1 to 3 after the IL1α injection. The analysis of EPCP engraftment demonstrated a significant and reproducible positive correlation between the 10panx peptide or Panx1 sdRNAi treatment and the number of engrafted cells. Similarly, treatment of the LG of the TSP-1-/- mouse (mouse model of chronic LG inflammation) by either Panx1 or Caspase-4 (also known as Casp11) sdRNAi showed a significant decrease in expression of proinflammatory markers and the lymphocyte infiltration.
Our results suggest that blocking Panx1 and/or Casp4 activities is a beneficial strategy to enhance donor cell engraftment and LG regeneration through the reduction of inflammation.
干燥综合征是一种全身性慢性自身免疫性炎症性疾病，主要累及唾液腺和泪腺（LGs）。目前尚无治愈方法；因此，基于细胞的再生疗法可能是一种可行的选择。细胞外ATP促进LG炎症，并由泛连接蛋白1（Panx1）膜通道糖蛋白介导。我们提出，通过操纵Panx1活性来抑制炎症可以刺激上皮细胞祖细胞（EPCP）植入。
通过qRT-PCR和免疫染色检测小鼠和人LG中泛连接蛋白的表达。通过注射白细胞介素-1α（IL1α）诱导急性LG炎症。在EPCP移植前，用Panx1特异性阻断肽（10panx）或可自我递送的RNA干扰（sdRNAi）处理血小板反应蛋白-1基因敲除小鼠（TSP-1-/-）的IL1α损伤或慢性炎症的LGs。通过显微镜分析细胞植入的效果和炎症区域。
在小鼠和人LGs中检测到Panx1和Panx2。在IL1α注射后第1至3天的急性炎症期间，Panx1和促炎因子上调。EPCP植入分析表明，10panx肽或Panx1 sdRNAi处理与植入细胞数量之间存在显著且可重复的正相关。同样，用Panx1或半胱天冬酶-4（也称为Casp11）sdRNAi处理TSP-1-/-小鼠（慢性LG炎症小鼠模型）的LG，促炎标志物的表达和淋巴细胞浸润显著减少。
我们的结果表明，通过减少炎症来阻断Panx1和/或Casp4活性是增强供体细胞植入和LG再生的有益策略。
