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Logic-gated tumor-microenvironment nanoamplifier enables targeted delivery of CRISPR/Cas9 for multimodal cancer therapy

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Acta Pharmaceutica Sinica B 2024年第2期14卷 795-807页

作者： Yongchun Pan Xiaowei Luan Fei Zeng Xuyuan Wang Shurong Qin Qianglan Lu Guanzhong he Yanfeng Gao Xiaolian Sun Xin Han bangshun he Yujun Song Department of Laboratory Medicine Nanjing First HospitalNanjing Medical UniversityNanjing 210006China College of Engineering and Applied Sciences State Key Laboratory of Analytical Chemistry for Life ScienceNanjing UniversityNanjing 210023China State Key Laboratory of Natural Medicines Key Laboratory of Drug Quality Control and PharmacovigilanceDepartment of Pharmaceutical AnalysisChina Pharmaceutical UniversityNanjing 211198China School of Medicine&Holistic Integrative Medicine Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources IndustrializationNanjing University of Chinese MedicineNanjing 210023China

Recent innovations in nanomaterials inspire abundant novel tumor-targeting CRISPR-based gene ***,the therapeutic efficiency of traditional targeted nanotherapeutic strategies is limited by that the biomarkers vary in a spatiotemporal-dependent manner with tumor ***,we propose a self-amplifying logic-gated gene editing strategy for gene/H_(2)O_(2)-mediated/starvation multimodal cancer *** this approach,a hypoxia-degradable covalent-organic framework(COF) is synthesized to coat a-ZIF-8 in which glucose oxidase(GOx) and CRISPR system are *** intensify intracellular redox dyshomeostasis,DNAzymes which can cleave catalase mRNA are loaded as *** the nano system gets into the tumor,the weakly acidic and hypoxic microenvironment degrades the ZIF-8@COF to activate GOx,which amplifies intracellular H^(+)and hypoxia,accelerating the nanocarrier degradation to guarantee available CRISPR plasmid and GOx release in target *** tandem reactions deplete glucose and oxygen,leading to logic-gated-triggered gene editing as well as synergistic gene/H_(2)O_(2)-mediated/starvation ***,this approach highlights the biocomputing-based CRISPR delivery and underscores the great potential of precise cancer therapy.

关键词： CRISPRdelivery Enzymeencapsulation Logic gate Hybridmaterial Hypoxia Gene editing Precise nanomedicine Multimodal therapy

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Photothermal probiotics delivery of photodynamic agents for enhanced immunogenic cell death within hypoxia center of solid tumors

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Nano Research 2024年第1期18卷 642-652页

作者： Wen Luo Shiyu Du Yateng Jiang bangshun he Jun Tang Jingjing Yang Yujun Song 1Jiangsu Key Laboratory of Artificial Functional Materials State Key Laboratory of Analytical Chemistry for Life Science National Laboratory of Solid State Microstructures College of Engineering and Applied Sciences Nanjing University Nanjing 210023 China 2Department of Laboratory Medicine Nanjing First Hospital Nanjing Medical University Nanjing 210006 China 3Department of Anesthesiology Jinling Hospital Medical School of Nanjing University Nanjing 210000 China 4Department of Biochemistry and Molecular Biology School of Medicine & Holistic Integrative Medicine Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization Nanjing University of Chinese Medicine Nanjing 210023 China

Resulting from harsh hypoxic environmental conditions, central cells in the core of solid tumors are usually more aggressive and malignant with a less stable genome. Therefore, therapeutic agents with improved penetration for the activation of immunity in tumor centers exhibit promising potential in immunotherapies. here, polydopamine-coated Escherichia coliNissle 1917 (EcN) bearing chlorin e6 (Ce6)-loaded and polyethyleneimine (PEI)-coated hollow manganese dioxide (shorted as EP-ChP) are applied for enhanced immunotherapy in deep tumors. After accumulation in tumor center through hypoxia targeting, manganese dioxide is degradated under the tumor microenvironment with released Ce6 and thus generates reactive oxygen species (ROS) upon 660 nm laser irradiation, which can further lower thermal-resistance of cancer cells via HSP90α downregulation. Owing to that, heating induced by polydopamine upon 808 nm laser irradiation can achieve effective tumor ablation. Phototherapy upon dual laser induces enhanced immunogenic cell death, while bacterial infections in tumor tissues also trigger innate immunity. This bacteria-based approach provides enhanced antitumor immune responses in deep tumors with great potential in the reshaping of immunosuppression tumor microenvironment.

关键词： bacteria phototherapies immunotherapy hypoxia targeting tumor penetration

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A nomogram based on serum bilirubin and albumin levels predicts survival in gastric cancer patients

A nomogram based on serum bilirubin and albumin levels predi...

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2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： Huiling Sun bangshun he Zhenlin Nie Yuqin Pan Kang Lin Shukui Wang General Clinical Research Center Nanjing First HospitalNanjing Medical University

Background Serum bilirubin, which is an end product of heme metabolism, was not thought to serve any physiological function. Nevertheless, recent studies have demonstrated that serum bilirubin has potent antioxidant, anti-inflammatory, and anticancer effects in colorectal cancer. The protective effects of serum bilirubin have been reported in lung cancer, colorectal cancer, and breast cancer. Additionally, decreased levels of albumin, a factor commonly used as an indicator of nutritional status, are associated with worse outcomes in gastric cancer *** We examined the predictive value of serum bilirubin and albumin levels in 778 gastric cancer patients from a single hospital in China who were divided among prospective training and retrospective validation cohorts. X-tile software was used to identify optimal cutoff values for separating training cohort patients into higher and lower overall survival(OS) groups, based on total bilirubin(TBIL) and albumin levels. Differences between high-and low-level group patients were evaluated using the χ2 test or Mann-Whitney U test. Survival analysis was performed using the Kaplan-Meier curve and log-rank test. Significant prognostic predictors from the survival analysis were included in a multivariate analysis using the Cox proportional hazards model. The nomogram for significant factors associated with 5-year overall survival(OS) was constructed using R software via a stepwise algorithm, and Harrell’s concordance index(c-index) was used to compare the performance of the nomogram to that of the TNM staging system. A calibration curve comparing observed outcomes to predicted outcomes was generated to further evaluate the nomogram’s accuracy in predicting *** Patients in the high albumin group were younger and more likely to have T1-T2, N0, or M0 stage disease than those in low albumin group. Similar results were obtained when validation cohort patients were divided into high and low level groups for eac

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MiR-216b functions as a tumor suppressor by targeting HMGB1 and inhibiting JAK2/STAT3 signaling way in colorectal cancer

MiR-216b functions as a tumor suppressor by targeting HMGB1 ...

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2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： Chen Xiaoxiang Liu Xiangxiang he bangshun Pan Yuqin Sun Huiling Shukui Wang Nanjing First Hospital Nanjing Medical University

Purpose:Mi R-216 b is implicated in the development of multiple types of cancers, however, a role for miR-216 b in colorectal cancer(CRC) remains elusive. The present study was performed to investigate the function and underlying mechanism of miR-216 b in human CRC tissues and cell lines. Methods : Quantitative RT-PCR(qRT-PCR) was employed to detect the expression of miR-216 b and HMGB1 mRNA in CRC tissues and cell lines. HMGB1 protein in CRC tissues and cell lines were detected by immunohistochemistry(IHC) and western blot analysis. The relationship between miR-216 b and its target gene HMGB1 were detected by bioinformatics methods and luciferase reporter assay. In addition, CCK-8, wound-healing and transwell assay were performed to analyze the biological functions of miR-216 b in CRC cells. Results:Mi R-216 b was significantly reduced in CRC tissues and cell lines compared with corresponding adjacent normal tissues(ANTs) and colonic mucosal epithelial cell line(FHC), and was markedly associated with the TNM stage, lymph node metastases, differentiation and poor over survival(OS)(phermore, the expression of HMGB1 was elevated and negatively correlated with miR-216 b levels in CRC. Additionally, Upregulation of mi R-216 b by transfecting with miR-216 b mimic significantly suppressed CRC cell proliferation, migration, invasion and decreased the expression of HMGB1. Finally, Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3) activity were found to be reduced by miR-216 b overexpression. Conclusions : Mi R-216 b might be a tumor suppressor by directly targeting HMGB1 and inhibiting JAK2/STAT3 signaling pathway in CRC.

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The inhibition role of miR-485-5p in colorectal cancer proliferation and invasion via targeting CD147

The inhibition role of miR-485-5p in colorectal cancer proli...

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2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： Yuqin Pan Huiling Sun Xiuxiu Hu bangshun he Xiangxiang Liu Shukui Wang Central Laboratory Nanjing First HospitalNanjing Medical University

Objective: The present study was designed to explore the expression and biological role of miR-485-5 p in the colorectal cancer(CRC). Methods: The expression of miR-485-5 p in forty-seven pairs of CRC and adjacent non-tumor tissues and CRC cell lines were detected by quantitative RT-PCR. CCK8 proliferation assay and invasion assay were performed to test the proliferation and invasion of CRC cells after overexpression of miR-485-5 p. The effect of miR-485-5 p on CRC in vivo was validated by murine xenograft model. The relationship of miR-485-5 p and its target gene CD147 was also investigated. Results: We found that the expression of miR-485-5 p in CRC tissues and cell lines were much lower than that in control, respectively. Moreover, the overexpression of miR-485-5 p could inhibit the proliferation and invasion of CRC cell lines in vitro and in vivo. Furthermore, we found that miR-485-5 p could decrease the expression of CD147 by directly targeting CD147 3’UTR. In addition, the expression of CD147 was inversely correlated with the expression of miR-485-5 p in CRC tissues. Conclusions: Our results suggested that miR-485-5 p play a pivotal role of tumor-suppressing in CRC through down-regulating cancer-associated gene CD147, which may provide a better understanding of the pathogenesis in CRC and a new bio-target for CRC therapy.

关键词： MiR-485-5p Tumor-suppressing Colorectal cancer CD147 Pathogenesis

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兔外周血检测早期内皮祖细胞方法的比较

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中华实验外科杂志 2014年第10期31卷 2161-2163页

作者：林爱翠周奕戈何帮顺张丽佳周末南京医科大学附属南京医院(南京市第一医院) 210006

目的对内皮祖细胞（EPCs）检测中常用的密度梯度离心法和两种裂解红细胞的方法进行比较,探索对血液中EPCs进行以鉴定数量、含量为目的的最简便、经济、准确的实验方法.方法取新西兰白兔动脉血存储于含肝素的真空采血管中,分别采用淋... 详细信息

目的对内皮祖细胞（EPCs）检测中常用的密度梯度离心法和两种裂解红细胞的方法进行比较,探索对血液中EPCs进行以鉴定数量、含量为目的的最简便、经济、准确的实验方法.方法取新西兰白兔动脉血存储于含肝素的真空采血管中,分别采用淋巴细胞分离液、流式细胞溶血素和红细胞裂解液对血液进行预处理.所获细胞采用抗小鼠藻蓝蛋白（APC）荧光素标记的CD3（CD3-APC）、抗小鼠藻红蛋白（PE）荧光素标记的CD45（CD45-PE）、抗小鼠异硫氰酸荧光素（FITC）标记的CD133 Prominin-1[CD133-（Prominin-1）-FITC]进行标记后通过流式细胞仪检测早期EPCs含量.结果 3种方法检测早期EPCs含量差异无统计学意义（P＞0.05）,其中流式细胞溶血素法所需的血液、处理液、试管和仪器最少,方法最简便,操作中影响实验结果的因素最少,有利于重复处理和测试.结论流式细胞溶血素法是检测外周血EPCs预处理方法中最容易实施的一种方法.

关键词：预处理早期内皮祖细胞裂解密度梯度离心

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叶酸代谢相关酶单核苷酸基因多态性与乳腺癌的相关性研究

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南京医科大学学报（自然科学版） 2016年第4期36卷 473-478页

作者：陶世龙王欢何帮顺徐勇朱锋南京市江宁区疾病预防控制中心江苏南京211100 南京医科大学附属江宁医院检验科江苏南京211100 南京医科大学附属南京医院中心实验室江苏南京210006

目的：探讨5个叶酸代谢相关酶基因多态性位点（MTRR,rs1801394;MTR,rs1805087;MTHFR,rs1801133;FHTFD,rs2002287;FTHFD,rs2276731）与中国女性乳腺癌发病风险及病理特征的相关性。方法：采用病例对照研究,选取298例女性乳腺癌患者为病... 详细信息

目的：探讨5个叶酸代谢相关酶基因多态性位点（MTRR,rs1801394;MTR,rs1805087;MTHFR,rs1801133;FHTFD,rs2002287;FTHFD,rs2276731）与中国女性乳腺癌发病风险及病理特征的相关性。方法：采用病例对照研究,选取298例女性乳腺癌患者为病例组及298例正常女性为对照组,采用Massarray技术检测5个多态性位点的基因型,采用Logistic回归法分析不同基因型与乳腺癌发生及病理特征的关系。结果：rs1801394位点的各基因型在2组中分布差异有统计学意义（χ^2=9.900,P=0.007）。AG基因型在乳腺癌组中的频率（28.52%）显著低于对照组（38.59%）。结论：MTRR rs1801394位点AG基因型与乳腺癌的发生相关,且受乳腺癌病理参数和肿瘤组织标本中雌激素受体（estrogen receptor,ER）、孕激素受体（progesterone receptor,PR）及原癌基因人类表皮生长因子受体2（human epidermalgrowth factor receptor-2,heR2,也称erb B-2）水平等因素影响。

关键词：乳腺癌叶酸代谢相关酶基因单核苷酸多态性

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miR-224在弥漫大B细胞淋巴瘤患者中的表达及其临床意义

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中华血液学杂志 2014年第7期35卷 619-622页

作者：宋国齐顾玲何帮顺潘玉琴王书奎东南大学医学院南京210009

目的分析miR-224在弥漫大B细胞淋巴瘤（DLBCL）患者中的表达及其与临床病理特征和预后的关系.方法以168例DLBCL患者为研究对象,以25份无肿瘤细胞侵犯的正常淋巴结组织标本为对照,采用Real-time PCR技术检测其miR-224的相对表达水平,... 详细信息

目的分析miR-224在弥漫大B细胞淋巴瘤（DLBCL）患者中的表达及其与临床病理特征和预后的关系.方法以168例DLBCL患者为研究对象,以25份无肿瘤细胞侵犯的正常淋巴结组织标本为对照,采用Real-time PCR技术检测其miR-224的相对表达水平,并探讨miR-224表达水平与患者临床病理特征及预后的关系.结果 DLBCL患者淋巴结组织中miR-224表达水平为0.97±0.33,对照组为1.87±0.43,两者比较差异有统计学意义（P＜0.05）.miR-224在DLBCL患者中的表达与患者年龄（P=0.434）、性别（P=0.613）、疾病分期（P=0.250）、国际预后指数（P=0.355）、LDH（P=0.398）均无相关性.以DLBCL患者miR-224表达水平的平均值为阈值,将患者分为高表达组和低表达组,两组患者的5年无进展生存率及总生存率差异均有统计学意义（P值均＜0.05）,前者均较后者高.结论 miR-224可能作为抑癌基因干扰DLBCL的发生、发展,其表达水平可作为DLBCL患者预后判断的标志.

关键词：淋巴瘤,大B细胞,弥漫性微RNAs 预后

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阿司匹林对人胃黏膜上皮细胞生长和occludin蛋白表达的影响及其机制研究

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胃肠病学 2011年第7期16卷 400-403页

作者：张弓羽张振玉姜宗丹何帮顺胡可伟南京医科大学附属南京第一医院消化科 210006 南京医科大学附属南京第一医院中心实验室

背景:研究显示ERK信号通路激活可促进细胞生长,上调紧密连接蛋白occludin表达,而阿司匹林可抑制ERK的磷酸化激活。目的:探讨阿司匹林对人胃黏膜上皮细胞生长和occludin蛋白表达的影响及其可能机制。方法:建立人胃黏膜上皮细胞株GES-1单... 详细信息

背景:研究显示ERK信号通路激活可促进细胞生长,上调紧密连接蛋白occludin表达,而阿司匹林可抑制ERK的磷酸化激活。目的:探讨阿司匹林对人胃黏膜上皮细胞生长和occludin蛋白表达的影响及其可能机制。方法:建立人胃黏膜上皮细胞株GES-1单层细胞模型,MTT法检测阿司匹林(5～20 mmol/L)和MEK抑制剂U0126(10～40μmol/L)对GES-1细胞的生长抑制作用。将GES-1细胞分为阿司匹林(8.78 mmol/L)组、U0126(14.91μmol/L)组、联合组(U0126+阿司匹林)和阴性对照组,倒置相差显微镜下观察细胞形态学变化,流式细胞术检测细胞凋亡情况,蛋白质印迹法检测p-ERK1/2、occludin蛋白表达。结果:阿司匹林和U0126均能剂量依赖性地抑制GES-1细胞生长,联合组细胞凋亡较两药单用更为显著,贴壁细胞数量减少更为明显,细胞变圆、悬浮。阿司匹林组、U0126组和联合组p-ERK1/2、occludin蛋白表达量均显著低于阴性对照组,U0126组和联合组降低更为明显。结论:阿司匹林可抑制人胃黏膜上皮细胞生长,下调occludin蛋白表达,其机制至少部分与抑制ERK信号通路激活有关。

关键词：阿司匹林胃黏膜上皮细胞 Occludin 信号传导细胞外信号调节MAP激酶类

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Association of polymorphisms of LTA+252 and IL-10 with gastric cancer risk in Chinese population

Association of polymorphisms of LTA+252 and IL-10 with gastr...

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2010’全国肿瘤分子标志及应用学术研讨会暨第五届中国中青年肿瘤专家论坛

作者： bangshun he,Yuqin Pan,Yongfei Xu,Chan Zhu,Lili Qu,Zhipeng Tang,Qian Bao,Fuliang Tian,Shukui Wang~* Central Laboratory,Nanjing First Hospital Affiliated to Nanjing Medical University,Nanjing 210006,China.

Background:Gastric cancer is one of the most common malignancies afflicting the Chinese *** association between Polymorphisms in LTA+252 and interleukin-10 genes and gastric cancer risk remain ***:In this study,we

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