重编程小鼠结肠癌微环境助力树突细胞的抗肿瘤免疫疗效 BMC Journal

　　 论文标题：Reprogramming the murine colon cancer microenvironment using lentivectors encoding shRNA against IL-10 as a component of a potent DC-based chemoimmunotherapy

　　 期刊：Journal of Experimental Clinical Cancer Research

　　 作者：Joanna Rossowska, Natalia Anger, Agnieszka Szczygiel, Jagoda Mierzejewska and Elzbieta Pajtasz-Piasecka

　　 发表时间：2018/06/28

　　 数字识别码：10.1186/s13046-018-0799-y

　　 原文链接：https://jeccr.biomedcentral.com/articles/10.1186/s13046-018-0799-y?utm_source=BMC_mailingutm_medium=Email_Internalutm_content=EvaFon-BMC-Journal_of_Experimental_and_Clinical_Cancer_Research-Oncology-Globalutm_campaign=AJH_AWA_JECCR_10th_Anniversary#Abs1

　　 微信链接：https://mp.weixin.qq.com/s/XyyI09LCOtv0MnQ9GBzOjA

Journal of Experimental Clinical Cancer Research在线发表了一篇论文Reprogramming the murine colon cancer microenvironment using lentivectors encoding shRNA against IL-10 as a component of a potent DC-based chemoimmunotherapy 。作者利用靶向IL-10编码shRNA的慢病毒载体（shIL10 LVs）对小鼠结肠癌的微环境进行重编程，从而使其在树突细胞介导的化学免疫抗肿瘤治疗中发挥重要作用。

　　 肿瘤微环境（TME）中存在大量免疫抑制因子，从而降低了癌症疫苗的有效性。本研究的主要目的是在小鼠结肠癌模型MC38中使用shIL10 LVs对其内部微环境进行重构，进而提高基于树突细胞（DC）的免疫疗法以及由环磷酰胺（CY）和DC组成的化学免疫疗法抗肿瘤的有效性。

　　 波兰科学院的Joanna Rossowska及其同事通过实时PCR和ELISA法在体外和体内分别检测了shIL10 LVs沉默IL-10表达的效果。

图1： IL-10在体外和体内的沉默效果

b: qPCR检测IL-10表达量

c: 转入LVs两天后的EGFP表达量变化趋势

e-j: shIL10–3 在肿瘤内的活性

图源：Joanna Rossowska 等

　　 他们使用流式细胞仪检查肿瘤内接种的慢病毒载体对MC38肿瘤微环境的影响。通过肿瘤生长受抑程度和局部及全身免疫应答的活化状态来评估疗效。

图2：免疫治疗(EXP.A)或化学免疫治疗(EXP.B)后对MC38肿瘤生长的影响

a, b:处理方案

c, d:治疗以后MC38肿瘤生长的速度

图3：免疫治疗(EXP.A)或化学免疫治疗(EXP.B)后对机体免疫应答的影响

f, g: 实验A、B处理后的脾细胞活性

　　 此外，他们还发现肿瘤内接种shIL10 LVs对肿瘤细胞和TME浸润细胞具有转化作用，使得IL-10分泌明显减少。

　　 连续三周应用shIL10 LVs后，肿瘤生长受到抑制，但同时使用shIL10 LVs和BMDC/TAg时，抗肿瘤效果并未增强。但当使用CY进行预处理后，MC38肠肿瘤生长明显受到抑制，同时TME中MDSCs和Treg明显减少，局部和全身的Th1抗肿瘤反应被有效激活。

　　 这篇研究的结果表明，shIL10 LV和CY能重塑TME，增强DC活性，并帮助DC细胞增殖，促进其抗肿瘤免疫反应。 因此，未来可以在化学免疫治疗的基础之上，进一步探究使用慢病毒载体消除局部IL-10的治疗方案。

　　 摘要：

Background

The excessive amounts of immunosuppressive factors present in a tumor microenvironment (TME) reduce the effectiveness of cancer vaccines. The main objective of our research was to improve the effectiveness of dendritic cell (DC)-based immunotherapy or chemoimmunotherapy composed of cyclophosphamide (CY) and DCs by application of lentivectors encoding shRNA specific to IL-10 (shIL10 LVs) in murine colon carcinoma MC38 model.

Methods

The efficacy of shIL10 LVs in silencing of IL-10 expression was measured both in vitro and in vivo using Real-Time PCR and ELISA assays. In addition, the influence of intratumorally inoculated lentivectors on MC38 tumor microenvironment was examined using flow cytometry method. The effect of applied therapeutic schemes was determined by measurement of tumor growth inhibition and activation state of local and systemic immune response.

Results

We observed that intratumorally inoculated shIL10 LVs transduced tumor and TME-infiltrating cells and reduced the secretion of IL-10. Application of shIL10 LVs for three consecutive weeks initiated tumor growth inhibition, whereas treatment with shIL10 LVs and BMDC/TAg did not enhance the antitumor effect. However, when pretreatment with CY was introduced to the proposed scheme, we noticed high MC38 tumor growth inhibition accompanied by reduction of MDSCs and Tregs in TME, as well as activation of potent local and systemic Th1-type antitumor response.

Conclusions

The obtained data shows that remodeling of TME by shIL10 LVs and CY enhances DC activity and supports them during regeneration and actuation of a potent antitumor response. Therefore, therapeutic strategies aimed at local IL-10 elimination using lentiviral vectors should be further investigated in context of combined chemoimmunotherapies.

　　 阅读论文全文请访问：https://jeccr.biomedcentral.com/articles/10.1186/s13046-018-0799-y?utm_source=BMC_mailingutm_medium=Email_Internalutm_content=EvaFon-BMC-Journal_of_Experimental_and_Clinical_Cancer_Research-Oncology-Globalutm_campaign=AJH_AWA_JECCR_10th_Anniversary#Abs1

　　 期刊介绍：Journal of Experimental Clinical Cancer Research (https://jeccr.biomedcentral.com/) is an online peer-reviewed journal that publishes original research papers, reviews and commentaries in cancer research, from bench to bedside.

　　 2017 Journal Metrics

　　 Citation Impact

　　 6.217 - 2-year Impact Factor

　　 5.280 - 5-year Impact Factor

　　 1.350 - Source Normalized Impact per Paper (SNIP)

　　 2.000 - SCImago Journal Rank (SJR)

　　 （来源：科学网）

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