CAR expression in invasive breast carcinoma and its effect on adenovirus transduction efficiency

IF 6.1 1区 医学 Q1 ONCOLOGY
Abraham T. Phung, Jaimin R. Shah, Tao Dong, Tony Reid, Christopher Larson, Ana B. Sanchez, Bryan Oronsky, William C. Trogler, Andrew C. Kummel, Omonigho Aisagbonhi, Sarah L. Blair
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引用次数: 0

Abstract

Breast cancer is the second leading cause of death in women, with invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) as the two most common forms of invasive breast cancer. While estrogen receptor positive (ER+) IDC and ILC are treated similarly, the multifocality of ILC presents challenges in detection and treatment, worsening long-term clinical outcomes in patients. With increasing documentation of chemoresistance in ILC, additional treatment options are needed. Oncolytic adenoviral therapy may be a promising option, but cancer cells must express the coxsackievirus & adenovirus receptor (CAR) for adenoviral therapy to be effective. The present study aims to evaluate the extent to which CAR expression is observed in ILC in comparison to IDC, and how the levels of CAR expression correlate with adenovirus transduction efficiency. The effect of liposome encapsulation on transduction efficiency is also assessed. To characterize CAR expression in invasive breast carcinoma, 36 formalin-fixed paraffin-embedded (FFPE) human breast tumor samples were assayed by CAR immunohistochemistry (IHC). Localization of CAR in comparison to other junctional proteins was performed using a multiplex immunofluorescence panel consisting of CAR, p120-catenin, and E-cadherin. ILC and IDC primary tumors and cell lines were transduced with E1- and E3-deleted adenovirus type 5 inserted with a GFP transgene (Ad-GFP) and DOTAP liposome encapsulated Ad-GFP (DfAd-GFP) at various multiplicities of infection (MOIs). Transduction efficiency was measured using a fluorescence plate reader. CAR expression in the human primary breast carcinomas and cell lines was also evaluated by IHC. We observed membranous CAR, p120-catenin and E-cadherin expression in IDC. In ILC, we observed cytoplasmic expression of CAR and p120-catenin, with absent E-cadherin. Adenovirus effectively transduced high-CAR IDC cell lines, at MOIs as low as 12.5. Ad-GFP showed similar transduction as DfAd-GFP in high-CAR IDC cell lines. Conversely, Ad-GFP transduction of ILC cell lines was observed only at MOIs of 50 and 100. Furthermore, Ad-GFP did not transduce CAR-negative IDC cell lines even at MOIs greater than 100. Liposome encapsulation (DfAd-GFP) improved transduction efficiency 4-fold in ILC and 17-fold in CAR-negative IDC cell lines. The present study demonstrates that oncolytic adenoviral therapy is less effective in ILC than IDC due to differences in spatial CAR expression. Liposome-enhanced delivery may be beneficial for patients with ILC and tumors with low or negative CAR expression to improve adenoviral therapeutic effectiveness.
浸润性乳腺癌中的 CAR 表达及其对腺病毒转导效率的影响
乳腺癌是女性的第二大死因,浸润性导管癌(IDC)和浸润性小叶癌(ILC)是两种最常见的浸润性乳腺癌。虽然雌激素受体阳性(ER+)的浸润性导管癌和浸润性小叶癌的治疗方法类似,但浸润性小叶癌的多发性给检测和治疗带来了挑战,使患者的长期临床预后恶化。随着ILC化疗耐药的文献越来越多,我们需要更多的治疗方案。肿瘤溶解性腺病毒疗法可能是一种很有前景的选择,但癌细胞必须表达柯萨奇病毒和腺病毒受体(CAR),腺病毒疗法才能有效。本研究旨在评估与IDC相比,CAR在ILC中的表达程度,以及CAR表达水平与腺病毒转导效率的相关性。本研究还评估了脂质体封装对转导效率的影响。为了确定CAR在浸润性乳腺癌中的表达特征,36份福尔马林固定石蜡包埋(FFPE)的人类乳腺肿瘤样本通过CAR免疫组化(IHC)进行了检测。使用由 CAR、p120-catenin 和 E-cadherin 组成的多重免疫荧光面板对 CAR 与其他交界蛋白的定位进行了比较。用插入 GFP 转基因(Ad-GFP)的 E1 和 E3 缺失的 5 型腺病毒和 DOTAP 脂质体包裹的 Ad-GFP(DfAd-GFP)以不同的感染倍数(MOIs)转导 ILC 和 IDC 原发肿瘤和细胞系。使用荧光平板阅读器测量转导效率。我们还通过 IHC 评估了 CAR 在人类原发性乳腺癌和细胞系中的表达。我们观察到 IDC 中膜上 CAR、p120-catenin 和 E-cadherin 的表达。在 ILC 中,我们观察到 CAR 和 p120-catenin 在细胞质中表达,而 E-cadherin 则缺失。腺病毒能有效地转导高CAR的IDC细胞系,MOI可低至12.5。在高CAR IDC细胞系中,Ad-GFP的转导与DfAd-GFP相似。相反,Ad-GFP 转导 ILC 细胞系的情况只有在 MOI 为 50 和 100 时才能观察到。此外,Ad-GFP 也不能转导 CAR 阴性的 IDC 细胞系,即使 MOI 超过 100。脂质体封装(DfAd-GFP)在 ILC 细胞系中的转导效率提高了 4 倍,在 CAR 阴性的 IDC 细胞系中提高了 17 倍。本研究表明,由于CAR空间表达的差异,溶瘤腺病毒疗法在ILC中的效果不如IDC。脂质体增强递送可能有利于ILC患者和CAR低表达或阴性的肿瘤患者,从而提高腺病毒的治疗效果。
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来源期刊
Breast Cancer Research
Breast Cancer Research 医学-肿瘤学
自引率
0.00%
发文量
76
期刊介绍: Breast Cancer Research is an international, peer-reviewed online journal, publishing original research, reviews, editorials and reports. Open access research articles of exceptional interest are published in all areas of biology and medicine relevant to breast cancer, including normal mammary gland biology, with special emphasis on the genetic, biochemical, and cellular basis of breast cancer. In addition to basic research, the journal publishes preclinical, translational and clinical studies with a biological basis, including Phase I and Phase II trials.
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