失调的祖细胞群可能导致胎盘发育不良和胎盘功能不全的发病机理。侧群细胞具有祖细胞特性。最近的人类滋养层侧群分离鉴定了8种特定基因的富集(CXCL8,ELL2,GATA6,HK2,HLA-DPB1,INTS6,SERPINE3和UPP1)(Gamage等人。2020年,干细胞RevRep)。我们对人类胎盘和胎盘功能不全疾病中的这些滋养层侧群标记进行了表征:先兆子痫和胎儿生长受限(FGR)。在早产控制中,滋养细胞侧种群标记位于胎盘绒毛基底膜内衬的单核滋养细胞,先兆子痫和FGR胎盘切片(n=3,3个标记的小组/连续切片)。分析类器官人类滋养层干细胞(hTSC)到绒毛外滋养层(EVT)分化模型的单细胞转录组学(Shannon等人2022年,开发)确定所有侧群基因都富集在单核滋养细胞和在hTSC培养条件下致力于分化的滋养细胞中。通过96小时时程hTSC分化为EVT或合胞滋养层(n=5)的体外验证表明ELL2和HK2随着分化而增加(分别为p<0.0024,p<0.0039)。CXCL8和HLA-DPB1下调(分别为p<0.030,p<0.011)。GATA6和INTS6仅随EVT分化而增加,UPP1随着融合而减少。SERPINE3检测不到。在人胎盘中测量滋养层侧群标记mRNA(<34周妊娠;n=78先兆子痫,n=30FGR,n=18个妊娠匹配的对照)。与对照组相比,先兆子痫(分别为p=0.0006,p<0.0001,p=0.0335)和FGR胎盘(分别为p=0.0065,p<0.0001,p=0.0001)中的ELL2,HK2和CXCL8升高。先兆子痫和FGR妊娠的胎盘GATA6减少(分别为p=0.0014,p=0.0146)。仅FGR降低胎盘INTS6(p<0.0001)。这项研究确定了富含侧群标记的独特滋养层亚群的定位。某些侧群标记物的异常表达可能表明胎盘功能不全中独特的滋养细胞亚型的破坏。
Dysregulated progenitor cell populations may contribute to poor placental development and placental insufficiency pathogenesis. Side-population cells possess progenitor properties. Recent human
trophoblast side-population isolation identified enrichment of 8 specific genes (CXCL8, ELL2, GATA6, HK2, HLA-DPB1, INTS6, SERPINE3 and UPP1) (Gamage et al. 2020, Stem Cell Rev Rep). We characterised these
trophoblast side-population markers in human placenta and in placental insufficiency disorders: preeclampsia and fetal growth restriction (FGR).
Trophoblast side-population markers localised to mononuclear trophoblasts lining the placental villous basement membrane in preterm control, preeclamptic and FGR placental sections (n = 3, panel of 3 markers/serial section). Analysis of single-cell transcriptomics of an organoid human trophoblast stem cell (hTSC) to extravillous trophoblast (EVT) differentiation model (Shannon et al. 2022, Development) identified that all side-population genes were enriched in mononuclear
trophoblast and trophoblasts committed to differentiation under hTSC culture conditions. In vitro validation via 96 h time course hTSC differentiation to EVTs or syncytiotrophoblasts (n = 5) demonstrated ELL2 and HK2 increased with differentiation (p < 0.0024, p < 0.0039 respectively). CXCL8 and HLA-DPB1 were downregulated (p < 0.030, p < 0.011 respectively). GATA6 and INTS6 increased with EVT differentiation only, and UPP1 reduced with syncytialisation. SERPINE3 was undetectable.
Trophoblast side-population marker mRNA was measured in human placentas (< 34-weeks\' gestation; n = 78 preeclampsia, n = 30 FGR, and n = 18 gestation-matched controls). ELL2, HK2 and CXCL8 were elevated in preeclamptic (p = 0.0006, p < 0.0001, p = 0.0335 respectively) and FGR placentas (p = 0.0065, p < 0.0001, p = 0.0001 respectively) versus controls. Placental GATA6 was reduced in pregnancies with preeclampsia and FGR (p = 0.0014, p = 0.0146 respectively). Placental INTS6 was reduced with FGR only (p < 0.0001). This study identified the localisation of a unique trophoblast subset enriched for side-population markers. Aberrant expression of some side-population markers may indicate disruptions to unique trophoblast subtypes in placental insufficiency.