Tumor-induced erythroid precursor-differentiated myeloid cells mediate immunosuppression and curtail anti-PD-1/PD-L1 treatment efficacy

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Central nervous system and peripheral cell labeling by vascular endothelial cadherin-driven lineage tracing in adult mice

Understanding the contribution of endothelial cells to the progenitor pools of adult tissues has the potential to inform therapies for human disease.To address whether endothelial cells transdifferentiate into non-vascular cell types,we performed cell lineage tracing analysis using transgenic mice engineered to express a fluorescent marker following activation by tamoxifen in vascular endothelial cadherin promoter-expressing cells(VEcad-CreERT2;B6 Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze).Activation of target-cell labeling following 1.5 months of ad libitum feeding with tamoxifen-laden chow in 4–5 month-old mice resulted in the tracing of central nervous system and peripheral cells that include:cerebellar granule neurons,ependymal cells,skeletal myocytes,pancreatic beta cells,pancreatic acinar cells,tubular cells in the renal cortex,duodenal crypt cells,ileal crypt cells,and hair follicle stem cells.As Nestin expression has been reported in a subset of endothelial cells,Nes-CreERT2 mice were also utilized in these conditions.The tracing of cells in adult Nes-CreERT2 mice revealed the labeling of canonical progeny cell types such as hippocampal and olfactory granule neurons as well as ependymal cells.Interestingly,Nestin tracing also labeled skeletal myocytes,ileal crypt cells,and sparsely marked cerebellar granule neurons.Our findings provide support for endothelial cells as active contributors to adult tissue progenitor pools.This information could be of particular significance for the intravenous delivery of therapeutics to downstream endothelial-derived cellular targets.The animal experiments were approved by the Boise State University Institute Animal Care and Use Committee(approval No.006-AC15-018)on October 31,2018.     

糖肾平对糖尿病肾病KKAy小鼠肾脏保护作用及对RhoA/ROCK信号通路影响的研究

目的：探讨糖肾平对糖尿病肾病（DiabeticKidneyDisease,DKD）KKAy小鼠肾保护作用及其对RhoA/ROCK信号通路的影响。方法：雌性10周龄SPF级KKAy小鼠60只,KK鼠料诱导10周建立DKD模型,随机分为模型组、厄贝沙坦组、糖肾平低（0.525g?kg-1）、中（1.05g?kg-1）、高（2.1g?kg-1）剂量组,10只雌性C57BL/6J小鼠为正常组。各治疗组灌胃给药,正常组、模型组灌胃等体积去离子水,每4周称体质量并测24h尿蛋白定量。第26周小鼠麻醉后摘眼球取血,称肾质量、测空腹血糖（FastingBloodGlucose,FBG）、血尿素氮（BloodUreaNitrogen,BUN）、血肌酐（Serumcreatinine,Scr）和甘油三酯（Triglyceride,TG）含量;HE、Mallory和PAS染色观察肾组织病理;免疫组化、原位杂交测肾组织转化生长因子-β1（Transforminggrowthfactor-β1,TGF-β1)、Ras同源基因家族成员A（RashomologgenefamilymemberA,RhoA）、Rho相关卷曲蛋白激酶（Rho-associatedcoiledcoil-containingproteinkinase1,ROCK1）、α-平滑肌肌动蛋白（α-SmoothMuscleActin,α-SMA）、E-钙黏素（ECadherin,E-Cad）mRNA及蛋白表达。结果：与模型组相比,各治疗组体质量、肾质量/体质量、尿蛋白降低,糖肾平中、高剂量组有显著性差异（P＜0.01）;肾病理损害明显减轻,FBG、BUN、Scr、TG含量降低（P＜0.01）,ECadherinmRNA及蛋白表达增加,TGF-β1、RhoA、ROCK1和α-SMAmRNA和蛋白表达减少,糖肾平中、高剂量组差异显著（P＜0.01）。结论：糖肾平对DKDKKAy小鼠肾的保护作用及抑制肾小管上皮细胞转分化的作用,可能与调节RhoA/ROCK信号通路有关。     

阻断Rac1对转化生长因子-β诱导视网膜色素上皮细胞行为改变的作用研究

目的 观察转化生长因子β1(TGF-β1)诱导视网膜色素上皮(RPE)细胞间质样转分化(EMT)过程中Rac1在其细胞学行为改变中的作用.方法 人RPE-19细胞分为空白组、TGF-β1组、TGF-β1+NSC23766组、NSC23766组.所有实验于加入TGF-β1前2h给予NSC23766以封闭Rac1受体.免疫荧光、蛋白免疫印迹法检测细胞中α-平滑肌肌动蛋白(α-SMA)表达;细胞划痕、侵袭及凝胶收缩实验,观察NSC23766对细胞迁徙、侵袭、凝胶收缩作用.结果 TGF-β1组细胞中α-SMA荧光表达较空白组、TGF-β1+ NSC23766增强,差异有统计学意义(F=825.314,P=0.003);细胞划痕实验结果显示,TGF-β1组细胞间距小于TGF-β1+ NSC23766组,差异有统计学意义(P24h=0.04,P48h =0.001);与空白组细胞间距比较,差异无统计学差异(P=0.278).细胞侵袭实验结果显示,TGF-β1组穿过纤维膜的细胞数与空白组、TGF-β1+ NSC23766组、NSC23766组穿过纤维膜的细胞数比较,差异无统计学意义(F=0.371,P=0.055).凝胶收缩实验结果显示,TGF-β1组能明显增加细胞的促凝胶收缩能力,组间差异有统计学意义(F=40.473,P=0.014),TGF-β1组相对TGF-β1+ NSC23766组,差异有统计学意义(P=0.022).结论 Rac1在TGF-β1诱导RPE细胞凝胶收缩改变中发挥作用;NSC23766可通过调控Rac1活化抑制RPE细胞学行为的改变.     

Malignant melanoma with metaplastic cartilaginous transdifferentiation: A case report

Malignant melanoma is notorious for its remarkable morphological variation and aberrant histopathological patterns. However, melanoma with prominent cartilaginous transdifferentiation simulating chondrosarcoma is extremely rare. A 75‐year‐old male developed a swelling in his left inguinal region and was diagnosed with a metastatic melanoma, which was found to harbor a BRAF V600E mutation. Later on, the left inguinal lymph node was excised and immunohistochemistry done on the specimen revealed an undifferentiated component negative for S‐100 protein, HMB‐45 and Melan‐A and a cartilaginous component positive for S‐100 protein and diffusely positive for BRAF V600E mutation. To our knowledge, there are around 14 cases reported in the literature of malignant melanoma with pure cartilaginous transdifferentiation. In all cases, immunohistochemistry of the cartilaginous component was positive for S‐100, which is not an indicator of melanoma because cartilage expresses S‐100. BRAF mutational studies support that the tumor arose from a common melanoma cell that harbored the mutation and subsequently transdifferentiated. This case illustrates the importance of an accurate and thorough clinical assessment when it comes to the diagnosis of melanomas as they are notable for their impressive degree of morphologic variability. Moreover, this report helps shed light on the use of immunohistochemical analysis to reach a definitive diagnosis.     

精原干细胞体外转化和转分化的研究进展

精原干细胞(SSC)在睾丸曲细精管内微环境的精确调控下,只能单向分化为精子。然而,SSC可在特定条件下转化为多能干细胞,甚至直接重编程转分化为功能性终末分化细胞。该特殊潜能使其在再生和精准医学领域具有很好的运用前景。     

脂肪来源干细胞体外成骨和成脂及成神经的诱导分化研究

目的 探讨脂肪组织来源干细胞 (adipose stem cell,ADSC) 的表面表型及向成骨、成脂和成神经多向分化的生物学特性.方法 取SD大鼠的腹股沟、附睾垫和腹膜后脂肪,Ⅰ型胶原酶消化、分离培养ADSC,检测第3代ADSC的CD29、CD44和CD45的表达情况.将ADSC加入成骨诱导剂,通过茜素红S染色鉴定成骨细胞;成脂诱导剂利用油红O染色鉴定脂肪细胞;神经干培养基诱导,免疫荧光染色鉴定诱导的神经元.结果 ADSC表达CD29(99.07±0.12)%、CD44(95.82±0.68)%和CD45(0.11±0.12)%.其经成骨诱导4周,茜素红S染色显示聚集的细胞团中央形成红色钙化结节,碱性磷酸酶染色可见到细胞的胞质内有紫红色颗粒;成脂诱导1周,油红O染色可见细胞质内有橙红色脂滴;经过神经干培养基诱导6 d后,免疫荧光染色显示诱导的细胞Nestin、NF-200及GFAP阳性表达.结论 ADSC表达间质干细胞的表面表型,体外经诱导培养后可向成骨细胞、脂肪细胞和神经元多向分化,表明 ADSC具有干细胞的多向分化潜能.