大脑类器官在胶质母细胞瘤中的应用进展

胶质母细胞瘤（GBM）是成人最常见的中枢神经系统恶性肿瘤，具有高度的异质性，瘤内不同细胞和肿瘤微环境之间存在复杂的相互作用。目前体外模型还不能准确再现脑肿瘤微环境，体内原位模型耗时长，成本高昂，且成功率不高。类器官作为一种新型的肿瘤研究模型能够高度模拟原位组织的生理结构和功能，可以稳定地维持肿瘤细胞在体内的特征，已被广泛应用于各种恶性肿瘤的研究。目前已经通过基因编辑技术诱导大脑类器官形成脑肿瘤，用以研究肿瘤发生的早期阶段，GBM干细胞与大脑类器官共培养可建立高通量药物筛选，直接培养建立GBM类器官可用来研究肿瘤的生物学特性及预测治疗反应。该文对大脑类器官进行简要介绍，回顾GBM类器官建立的不同实验技术方法，并对GBM类器官的应用进行了归纳。     

小胶质细胞/巨噬细胞在胶质母细胞瘤中作用的研究进展

胶质母细胞瘤（GBM）是成人最常见的颅内恶性肿瘤,具有极强的侵袭性。此外,由于化疗药物难以通过血脑屏障和胶质母细胞瘤的耐药性,及对放疗敏感性较差等特点,故预后极差。其中肿瘤微环境的改变起到了至关重要的作用,在微环境中胶质瘤相关的小胶质细胞/巨噬细胞（GAMs）的作用正逐渐被重视。GAMs不仅有中枢系统的常驻小胶质细胞,还有来自外周的巨噬细胞。GAMs还有截然不同的两种极化类型,即抑制肿瘤生长的M1表型和促进肿瘤生长的M2表型。并且GAMs不单单和肿瘤细胞具有联系,还与微环境中其他非癌性脑细胞也有互动。该文将从GAMs的来源、极化、与肿瘤微环境中各种细胞间的相互影响阐述其在GBM中的作用,并且从靶向治疗的角度探讨其最新的研究进展。     

胶质母细胞瘤分子靶向治疗研究进展

胶质母细胞瘤(glioblastoma,GBM)的分子靶向治疗是目前研究的热点。大量针对GBM进行分子靶向治疗的临床研究相继开展,部分靶向药物如贝伐单抗、西地尼布、西仑吉肽、恩扎妥林和尼妥珠单抗已完成了Ⅲ期临床试验。大部分Ⅲ期临床试验未得出预想的阳性结果 ,然而,研究结果也显示某些特定的亚型能从靶向治疗中获益。未来的研究应充分考虑到GBM的异质性,转变试图以一种靶向药物治疗所有GBM的观念。寻找能预测特定靶向治疗效果的分子标记物,在基因和分子分型的基础上进行个体化的精准靶向治疗。     

肿瘤治疗电场联合替莫唑胺化疗与单独使用替莫唑胺化疗对胶质母细胞瘤临床疗效比较的Meta分析

目的 探讨单独使用替莫唑胺（TMZ）与替莫唑胺联合肿瘤治疗电场（TTF）治疗胶质母细胞瘤安全性和疗效的比较。方法 检索Pubmed、Cochrance、Embase、Ovid、Scopus、Web of Science、中国知网、万方数据知识服务平台、维普中文期刊数据库、中国生物医学文献服务系统数据库、谷歌学术自建库至2020年4月5日的文献,筛选TMZ和TTF+TMZ进行疗效比较的随机对照研究,按照纳入和排除标准,把总体生存率（OS）和无进展生存期（PFS）作为结局指标,最后使用Review Manager进行统计分析。结果 最终纳入4篇研究,共1091例患者,其中单纯TMZ组381例,TTF+TMZ组710例。TTF+TMZ组的平均OS （26.9个月）和平均PFS （14.7个月）,优于单纯TMZ组的平均OS （12.63个月）和平均PFS （5个月）（P<0.01）。结论 TTF+TMZ治疗GBM的有效性优于单纯使用TMZ的患者。     

H19在多形性胶质母细胞瘤中的表达及对细胞系增殖、侵袭的影响

目的探讨长链非编码RNA(Long noncoding RNAs,LncRNAs)H19在人多形性胶质母细胞瘤((glioblastoma mutliforme,GBM)中的表达及其对U87-GBM生物学功能的影响及作用机理。方法实时荧光定量核酸扩增检测系统(Realtime Quantitative PCR Detecting System,qPCR)检测H19在GBM表达;CCK-8试剂盒(Cell Counting Kit-8,CCK-8)检测U87细胞增殖;流式细胞技术(Flow Cytometry,FCM)检测U87细胞凋亡及细胞周期;侵袭小室(transwell)检测U87细胞迁移和侵袭.蛋白质印迹法(Western blot)法检测P53表达。结果 H19在GBM中高表达,上调H19促进GBM的增殖、侵袭、迁移;抑制凋亡;增加了细胞周期S期比例;抑制了P53的表达。结论 H19促进GBM的致癌性。     

新型肿瘤电场治疗仪治疗复发脑胶质母细胞瘤的前瞻性、单中心、单臂探索性临床试验方案

多形性胶质母细胞瘤（GBM）是原发性脑肿瘤中恶性程度最高的一种肿瘤,不但进展迅速且预后极差。肿瘤电场治疗（TTF）已被批准用于治疗GBM且具有良好的疗效。但是,尚无有关中国GBM患者采用TTF治疗的临床资料。该文报告一项复发性GMB的前瞻性、单中心、单臂探索性临床试验方案。该试验采用笔者开发的新型肿瘤电场治疗系统ASCLU-300,对参加者进行治疗。主要是评价该治疗系统的安全性和可行性。通过评估与治疗相关不良反应的患者人数,及根据进展时间（TTP）和总生存期（OS）来评价治疗反应;根据神经肿瘤治疗反应评价(RANO)标准进行评估,通过对比MRI检查确定TTP,并通过随访收集OS数据;与治疗相关的不良反应评估将基于描述性方法;通过Kaplan-Meier方法评估TTP和OS。这项初步研究将为进一步的大规模试验打下坚实基础,同时笔者希望新型肿瘤电场治疗系统能够有效治疗GBM,为中国GBM患者提供新的治疗方案。（临床试验注册：NCT0441793;于2020年6月8日注册。）     

circRNA在胶质母细胞瘤中的研究进展

<正>胶质母细胞瘤（glioblastoma multiforme,GBM）是一种高度恶性的颅内原发性肿瘤，WHO分级Ⅳ级。目前，GBM的标准治疗是手术切除联合放化疗，但中位生存期仅为15个月，5年生存率不足5%。如何更好地治疗GBM，改善病人生存时间和生活质量仍然是一个巨大的挑战[1]。因此，研究GBM恶性进展的调控机制、探索GBM早期肿瘤标志物，对GBM的早期诊断、治疗和预后评估具有重要意义。环状RNA(circRNA)是一类单链共价闭合的非编码RNA，没有5’帽结构和3’末端多聚（A）尾巴。     

解读《复发性进展性胶质母细胞瘤的治疗指南》

胶质母细胞瘤（glioblastoma,GBM）是最常见的成人原发性脑肿瘤,年发病率约为5/10万[1]。即使采用手术、放疗和化疗等综合治疗,90%以上的GBM患者会出现复发或进展。目前,对复发性或进展性GBM尚缺乏标准治疗策略。为此,加拿大GBM委员会的多学科工作组在2007年制定GBM治疗指南的基础上,参照美国临床肿瘤协会制定的证据级别和推荐分级标准[2],最近又制定了《复发性/进展性GBM的治疗指南》,现解读如下,供临床参考。     

丙戊酸钠治疗小儿全面强直阵挛性发作相关认知功能障碍的影响因素分析

目的 探讨丙戊酸钠治疗全面强直阵挛性发作相关认知功能障碍的影响因素。方法 选取2017年1月至2019年5月在我院就诊的80例小儿全面强直阵挛性发作癫痫患者作为研究对象。所有患儿均给予丙戊酸钠治疗,后依据韦氏儿童智力量表（WlSC-CR）智商得分分为认知正常组（≥ 80分）和认知障碍组（<80分）。分析丙戊酸钠治疗3个月后认知功能障碍的影响因素。结果 两组患儿的发病年龄、每月发病频率、治疗剂量及疗程比较,差异有统计学意义（P<0.05）。回归分析显示,发病年龄<5岁、每月发病频率≥ 3次、治疗剂量≥ 35 mg/（kg·d）、治疗疗程≥ 6个月是丙戊酸钠治疗小儿癫痫后认知功能障碍的危险因素（P<0.05）。认知障碍组治疗3个月后每月发病频率、治疗剂量、治疗疗程与VIQ、PIQ、FIQ呈负相关（P<0.05）;发病年龄与VIQ、PIQ、FIQ呈正相关（P<0.05）。结论 发病年龄<5岁、每月发病频次≥ 3次、治疗剂量≥ 35 mg/（kg·d）、治疗疗程≥ 6个月是丙戊酸钠治疗小儿癫痫后认知功能障碍的危险因素。     

AEBP1在胶质母细胞瘤中的表达及临床意义

目的 探讨脂肪细胞增强剂结合蛋白1（AEBP1）在胶质母细胞瘤（GBM）中的表达及临床意义。方法 采用生物信息学分析方法,利用ONCOMINE、GEPIA和STRING数据库评估AEBP1在GBM中的表达情况及其预后价值。结果 AEBP1在GBM组织中表达水平显著高于正常组织。生存分析表明,AEBP1表达水平与GBM总体生存率呈显著负相关。KEGG分析显示与AEBP1紧密相关的蛋白质主要参与肿瘤转录失调以及局部粘连信号通路。结论 我们应用生物信息学分析显示AEBP1可能参与GBM发生、发展,AEBP1表达水平增高可能是诊断GBM的重要指标,并有望成为预后指标。然而,AEBP1在GBM中的具体作用机制及其作为预后评估和治疗靶点的价值有待进一步研究     

CDK4/6抑制剂在治疗脑胶质母细胞瘤中的研究进展

细胞周期素依赖性蛋白激酶在临床上已用于乳腺癌的治疗,其机制是促使抑癌基因Rb磷酸化,使细胞从G1期进入S期。CDK4/6抑制剂可以抑制乳腺癌、头颈癌、非小细胞肺癌等多种恶性肿瘤的活性。胶质母细胞瘤具有复发率高、生存期短、预后差等特点,治疗难度大,而CDK4/6理论上作为胶质母细胞瘤(GBM)治疗的靶点,CDK4/6抑制剂作用于cyclin D-CDK4/6-INK4-Rb通路,也同样可以抑制胶质母细胞瘤。作为有希望治疗胶质母细胞瘤药物,CDK4/6抑制剂在动物实验,肿瘤细胞系等研究中展示了大量的证据证明其对胶质母细胞瘤能产生抑制作用。同时CDK4/6抑制剂其研发方向是寻找预测标志物,探索联合治疗,以试图解决神经胶质母细胞瘤治疗中的耐药性和复发性中的关键问题,为治疗神经胶质瘤的研究提供新方向和思路。     

Rhabdoid glioblastoma: Case report and literature review

Rhabdoid glioblastoma is a recently described entity in which a glioblastoma is associated with a rhabdoid component. Although rhabdoid glioblastoma has not appeared in the new World Health Organization classification of tumors of the CNS, it has a specific morphological feature and highly aggressive clinic process. Up to now, there have been six cases of rhabdoid glioblastoma reported in the literature. We report rhabdoid glioblastoma in the right front temporal lobe from a 31‐year‐old Chinese man. This tumor consisted of rhabdoid tumor cells with an eccentric nucleus and an eosinophilic cytoplasm. The tumor had an area appearing to be glioblastoma with microvascular proliferation and necrosis, and lacked a primitive neuroectodermal tumor component, and a mesenchymal component. Vimentin, epithelial membrane antigen, GFAP and integrase interactor (INI‐1) expression were found in the tumor cells. Genetic abnormalities which include monosomy or a deletion of chromosome 22 were not found in this tumor. After 3 months post‐surgery, the tumor was widespread in leptomeningia and the patient died. In conclusion, rhabdoid glioblastoma is a rare glioblastoma with poor prognosis; the differential diagnosis contained other rhabdoid tumors. This case will contribute to the profile of rhabdoid glioblastoma with typical morphology and immunophenotype, genetic and clinic features.     

人胶质瘤干细胞内在自我更新能力

目的 了解胶质瘤干细胞内在的自我更新和增殖能力。方法 观察原代胶质瘤细胞在单纯改良Eagle/F12培养液（DMEM/F12）中胶质瘤干细胞球的形成,并检测其CD133、胶质纤维酸性蛋白（GFAP）、微管相关蛋白（MAP2）、髓磷脂碱性蛋白（MBP）的表达。通过二代球体形成、细胞增殖测定、分化实验分析其自我更新、增殖、多能分化能力。通过裸鼠移植瘤实验观察所分离细胞球细胞与原代培养胶质瘤细胞成瘤能力的差异。结果 在单纯DMEM/F12培养液中形成的胶质瘤细胞球细胞表达神经干细胞标记CD133,不表达分化标志GFAP、MAP2,少数细胞表达MBP。分离出的胶质瘤细胞球细胞可在单纯DMEM/F12培养基中增殖,并能形成CD133阳性的二代细胞球,可分化为GFAP、MAP2、MBP阳性表达的肿瘤细胞。裸鼠成瘤实验显示其成瘤能力显著高于原代胶质瘤细胞。结论 胶质瘤干细胞能在无血清、无外源性细胞因子培养基中形成肿瘤干细胞球,胶质瘤干细胞的自我更新和增殖不依赖于外源性生长因子,它可能拥有自我更新的自身活化机制。     

Epidermal growth factor receptor in glioblastoma

We compiled the current state of knowledge about the epidermal growth factor receptor (EGFR) in glioblastoma. Glioblastoma is one of the most common primary brain tumours and has an unfavourable prognosis despite aggressive treatment. These factors stimulate new research trials and a recent area of interest of neuro-oncologists is EGFR. This molecule is frequently altered in glioblastoma and constitutes the potential target for therapy. We decided to review the literature on biological structure of that molecule, its biological activity and the role in GBL with potential targeting it in the future neuro-oncological practice.     

Functional activation of EphA5 receptor does not promote cell proliferation in the aberrant EphA5 expressing human glioblastoma U-118 MG cell line

Eph receptors are a subfamily of receptor tyrosine kinases (RTKs), that are activated by ephrin ligands and appear to play important roles in axon guidance and cell migration during development of the nervous system. Over-expression or constitutive activation of Eph receptors has been linked with increased proliferation in various tumours. We have recently described lineage aberrant expression of EphA5 in primary human astrocytomas, glioblastomas and in the human glioblastoma U-118 MG cell line. A role for EphA5 expression in these tumours is not apparent, and we have investigated the cellular effects of EphA5 activation using the human glioblastoma U-118 MG cell line as a model. Immunofluorescent staining demonstrated cell surface expression of EphA5. Activation of the EphA5 receptor using an ephrin-A1 recombinant fusion protein resulted in tyrosine phosphorylation of EphA5 in a time-dependent manner. Exposure of U-118 MG glioblastoma cells to ephrin-A1 did not result in significant spontaneous or FCS-stimulated cell proliferation, though a marginal decrease was observed. This is in converse to the effects of Eph activation in other tumour cell lines, and is the first study to investigate EphA5 in glioblastoma cell lines.     

Transdifferentiation of glioblastoma stem-like cells into mural cells drives vasculogenic mimicry in glioblastomas

Recent evidence has shown that glioblastoma stem-like cells (GSCs) can transdifferentiate into endothelial cells and vascular-like tumor cells. The latter pattern of vascularization indicates an alternative microvascular circulation known as vasculogenic mimicry (VM). However, it remains to be clarified how the GSC-driven VM makes a significant contribution to tumor vasculature. Here, we investigated 11 cases of glioblastomas and found that most of them consisted of blood-perfused vascular channels that coexpress mural cell markers smooth muscle α-actin and platelet-derived growth factor receptor β, epidermal growth factor receptor, and vascular endothelial growth factor receptor 2 (Flk-1), but not CD31 or VE-cadherin. This microvasculature coexisted with endothelial cell-associated vessels. GSCs derived from patients with glioblastomas developed vigorous mural cell-associated vascular channels but few endothelial cell vessels in orthotopic animal models. Suppression of Flk-1 activity and gene expression abrogated GSC transdifferentiation and vascularization in vitro, and inhibited VM in animal models. This study establishes mural-like tumor cells differentiated from GSCs as a significant contributor to microvasculature of glioblastoma and points to Flk-1 as a potential target for therapeutic intervention that could complement current anti-angiogenic treatment.     

Human glioblastoma cell line 86HG39 activates T cells in an antigen specific major histocompatibility complex class II-dependent manner.

The capacity of three different human glioblastoma cell lines to activate human T cells was analysed by measuring major histocompatibility complex (MHC) antigen expression, monokine secretion and lectin, mAb OKT3 and antigen-driven T cell proliferation. All glioblastoma cells tested were able to induce PHA and concanavalin A (ConA)-driven T cell proliferation in a dose-dependent fashion, while all failed to induce T cell activation with mAb OKT3. In addition, the glioblastoma cell line 86HG39 was able to induce tetanus toxoid and toxoplasma lysate antigen-specific T cell proliferation. The responding T cell lines originated from only one out of five different donors. This foreign antigen-specific T cell proliferation induced by 86HG39 cells could be inhibited with mAb L243 directed against HLA-DR molecules. The study of monokine secretion by 86HG39 cells showed a strong interleukin (IL)-6 secretion after lipopolysaccharide (LPS) treatment, whilst no IL-1 secretion was observed. Furthermore, only 86HG39 cells were positive for HLA-DR molecules, whereas interferon (IFN) gamma treatment of 87HG28 and 87HG31 cells was necessary for the induction of class II antigen expression. Thus, cell line 86HG39 shows many features of an antigen presenting cell and the interaction of these cells with MHC compatible human T cells might be a useful model to study cellular immune reactions within the central nervous system.     

Amplification of epidermal growth factor receptor gene in gliomas: histopathology and prognosis.

In order to evaluate the incidence and prognostic significance of gene amplification in primary brain neoplasms we measured the number of gene copies per cell of three oncogenes (epidermal growth factor receptor [EGFR] gene, N-myc, C-myc) and syntenic control genes in 40 specimens using quantitative DNA dot blots. We observed EGFR gene amplification in astrocytomas and anaplastic astrocytomas with approximately the same incidence as in glioblastoma multiforme (33%), although large amplifications were only seen in glioblastoma multiforme. Fourteen patients had a supratentorial glioblastoma multiforme; six had EGFR gene amplification and eight had either normal EGFR gene copy number or elevated EGFR copy number attributable to extra copies of chromosome 7. Patients with gene amplification had shorter survival than patients without gene amplification (p = 0.01). The observed difference in survival was not likely to be due to group differences in age, sex, treatment, or histopathology.