CAR-T Cell Therapy for Breast Cancer: From Basic Research to Clinical Application

Breast cancer rises as the most commonly diagnosed cancer in 2020. Among women, breast cancer ranks first in both cancer incidence rate and mortality. Treatment resistance developed from the current clinical therapies limits the efficacy of therapeutic outcomes, thus new treatment approaches are urgently needed. Chimeric antigen receptor (CAR) T cell therapy is a type of immunotherapy developed from adoptive T cell transfer, which typically uses patients'' own immune cells to combat cancer. CAR-T cells are armed with specific antibodies to recognize antigens in self-tumor cells thus eliciting cytotoxic effects. In recent years, CAR-T cell therapy has achieved remarkable successes in treating hematologic malignancies; however, the therapeutic effects in solid tumors are not up to expectations including breast cancer. This review aims to discuss the development of CAR-T cell therapy in breast cancer from preclinical studies to ongoing clinical trials. Specifically, we summarize tumor-associated antigens in breast cancer, ongoing clinical trials, obstacles interfering with the therapeutic effects of CAR-T cell therapy, and discuss potential strategies to improve treatment efficacy. Overall, we hope our review provides a landscape view of recent progress for CAR-T cell therapy in breast cancer and ignites interest for further research directions.     

Characterization of the relationship between FLI1 and immune infiltrate level in tumour immune microenvironment for breast cancer

Breast cancer is the most common cancer and the leading cause of cancer death among women in the world. Tumour-infiltrating lymphocytes were defined as the white blood cells left in the vasculature and localized in tumours. Recently, tumour-infiltrating lymphocytes were found to be associated with good prognosis and response to immunotherapy in tumours. In this study, to examine the influence of FLI1 in immune system in breast cancer, we interrogated the relationship between the FLI1 expression levels with infiltration levels of 28 immune cell types. By splitting the breast cancer samples into high and low expression FLI1 subtypes, we found that the high expression FLI1 subtype was enriched in many immune cell types, and the up-regulated differentially expressed genes between them were enriched in immune system processes, immune-related KEGG pathways and biological processes. In addition, many important immune-related features were found to be positively correlated with the FLI1 expression level. Furthermore, we found that the FLI1 was correlated with the immune-related genes. Our findings may provide useful help for recognizing the relationship between tumour immune microenvironment and FLI1, and may unravel clinical outcomes and immunotherapy utility for FLI1 in breast cancer.     

HER-2抗独特型单克隆抗体疫苗-肿瘤疫苗的新探索

乳腺癌作为女性最常见的恶性肿瘤之一,发病率正呈逐年上升趋势。全球每13分钟就有一人死于乳腺癌,并随着发病的不断增加,死亡率也在明显上升。乳腺癌已经在女性恶性肿瘤发病率中占第一位,成为威胁女性健康的”头号杀手”。这其中,大部分的乳腺癌类型均为HER-2＋。许多的研究证明如果前期对HER-2过度表达的肿瘤应用针对HER-2的免疫疗法可以有效的控制肿瘤的发展。因此,建立体内的免疫应答可以控制过度表达HER-2的乳腺肿瘤病人的病情发展。随着曲妥珠单抗的产生和投入使用。启发人们发现更广阔的空间去构建针对HER-2的免疫应答的疫苗,从而使抗肿瘤的免疫作用更稳定更强大并且不会产生免疫耐受,进而防止肿瘤复发。因为现在针对HER-2产生的免疫耐受已经制约了肿瘤疫苗的发展,因此现在的研究热点是如何打破免疫耐受更好地发挥抗肿瘤的免疫作用。在这篇文章里,我们将着重介绍模拟HER-2的抗独特型单克隆抗体的应用。     

Melatonin is an appropriate candidate for breast cancer treatment: Based on known molecular mechanisms

Breast cancer is the most prevalent cancer and one of the most important causes of death in women throughout the world. Breast cancer risk factors include smoking, alcohol consumption, personal and family history, hypertension, and hormone therapy, long-term use of nonsteroidal anti-inflammatory drugs and tobacco usage. Surgery, chemotherapy, radiotherapy, immunotherapy, and neoadjuvant therapy are the current means for breast cancer treatment. Despite hormonal agents and chemotherapy, which have beneficial effects on lowering breast cancer death rate, the reaction of different people to these treatments is still a challenging point. Melatonin (N-acetyl-5-methoxy tryptamine) is a methoxy indole compound that is mainly secreted by the pineal gland at night; it is as an antioxidant, anti-inflammatory, and oncostatic agent. On the basis of recent studies, melatonin has antitumor properties on different cancer types and it may suppress cancer development in vitro and as well as in animal models. It is suggested that melatonin inhibits the development of breast cancer by various mechanisms. This paper summarizes the roles of melatonin in breast cancer treatment from the aspect of its molecular actions.     

The viral approach to breast cancer immunotherapy

Despite years of intensive research, breast cancer remains the leading cause of death in women worldwide. New technologies including oncolytic virus therapies, virus, and phage display are among the most powerful and advanced methods that have emerged in recent years with potential applications in cancer prevention and treatment. Oncolytic virus therapy is an interesting strategy for cancer treatment. Presently, a number of viruses from different virus families are under laboratory and clinical investigation as oncolytic therapeutics. Oncolytic viruses (OVs) have been shown to be able to induce and initiate a systemic antitumor immune response. The possibility of application of a multimodal therapy using a combination of the OV therapy with immune checkpoint inhibitors and cancer antigen vaccination holds a great promise in the future of cancer immunotherapy. Display of immunologic peptides on bacterial viruses (bacteriophages) is also increasingly being considered as a new and strong cancer vaccine delivery strategy. In phage display immunotherapy, a peptide or protein antigen is presented by genetic fusions to the phage coat proteins, and the phage construct formulation acts as a protective or preventive vaccine against cancer. In our laboratory, we have recently tested a few peptides (E75, AE37, and GP2) derived from HER2/neu proto-oncogene as vaccine delivery modalities for the treatment of TUBO breast cancer xenograft tumors of BALB/c mice. Here, in this paper, we discuss the latest advancements in the applications of OVs and bacterial viruses display systems as new and advanced modalities in cancer immune therapeutics.     

PD-1/PD-L1 pathway: Basic biology and role in cancer immunotherapy

Over the course of past few years, cancer immunotherapy has been accompanied with promising results. However, preliminary investigations with respect to immunotherapy concentrated mostly on targeting the immune checkpoints, nowadays, emerge as the most efficient strategy to raise beneficial antitumor immune responses. Programmed cell death protein 1 (PD-1) plays an important role in subsiding immune responses and promoting self-tolerance through suppressing the activity of T cells and promoting differentiation of regulatory T cells. PD-1 is considered as an immune checkpoint and protects against autoimmune responses through both induction of apoptosis in antigen-specific T cells and inhibiting apoptosis in regulatory T cells. Several clinical trials exerting PD-1 monoclonal antibodies as well as other immune-checkpoint blockades have had prosperous outcomes and opened new horizons in tumor immunotherapy. Nonetheless, a bulk of patients have failed to respond to these newly emerging immune-based approach and the survival rate was not satisfying. Additional strategies, especially combination therapies, has been initiated and been further promising. Attempts to identify novel and well-suited predictive biomarkers are also sensed. In this review, the promotion of cancer immunotherapy targeting PD-1 immunoinhibitory pathway is discussed.     

单细胞测序技术及其在乳腺癌研究中的应用

乳腺癌是起源于乳腺各级导管和乳腺上皮细胞,由增生到不典型增生而逐步发展成原位癌、早期浸润癌至浸润性癌的一种恶性肿瘤。传统高通量测序技术对乳腺癌的研究主要是鉴定与乳腺癌发生发展相关的"驱动基因",但是对于乳腺癌基因组结构变化以及亚克隆的鉴定等存在一定的局限性,并且忽略了乳腺癌肿瘤细胞之间的异质性。近年来兴起的单细胞测序技术是以单个细胞为研究对象,对基因拷贝和基因表达等数据进行分析,探究乳腺癌的细胞组成、细胞状态和细胞命运,绘制乳腺癌生态系统图谱,对临床患者进行精准分层,为实现个体化治疗提供支持。同时,还可以揭示乳腺癌与T细胞、巨噬细胞等免疫细胞间的相关性,为发现乳腺癌新的治疗靶点、免疫检查点等提供参考。本文对单细胞测序技术及其在乳腺癌研究中的应用和研究进展进行了综述,以期为单细胞测序技术的进一步发展提供参考,同时为理解乳腺癌的发病机制和免疫治疗提供基础支持。     

California's male suicide rate at record high.

Breast cancer is the most commonly occurring cancer in women and, until recently surpassed by lung cancer, was the leading cause of cancer-related death in women. It is the leading cause of death in women aged 39 to 44 years. The American Cancer Society has estimated that there will be 135,000 new cases of breast cancer and 42,300 breast cancer-related deaths in 1988. It is now predicted that breast cancer will develop in one out of every ten women in the United States. Given the clinical and public health significance of breast cancer, annual screening with mammography and clinical breast examination is recommended for women aged 50 and older to reduce breast cancer mortality.     

Therapeutic gene modified cell based cancer vaccines

History of cancer immunotherapy lasts for more than 120 years. In 1891 William B. Coley injected bacteria into inoperable cancer (bone sarcoma) and observed tumor shrinkage. He is recognized as the "'"Father of Immunotherapy"'". Cancer immunotherapy is based on the ability of the immune system to recognize cancer cells and to affect their growth and expansion. Beside the fact that, tumor cells are genetically distinct from their normal counterparts, and should be recognized and eliminated by immune system, the tumor associated antigens (TAAs) are often poorly immunogenic due to immunoediting. This process allows tumor to evolve during continuous interactions with the host immune system, and eventually escape from immune surveillance. Furthermore, tumor microenvironment consists of immunosuppressive cells that release immunosuppressive factors including IL-6, IL-10, IDO, TGFβ or VEGF. Interactions between cancer and stroma cells create network of immunosuppressive pathways, while activation of immune defense is inhibited. A key to successful immunotherapy is to overcome the local immunosuppression within tumor microenvironment and activate mechanisms that lead to tumor eradication. There are two clinical approaches of immunotherapy: active and passive. Active immunotherapy involves stimulation of immune response to tumor associated antigens (TAAs), either non-specifically via immunomodulating agents or specifically employing cancer vaccines. This review presents the progress and breakthroughs in design, development and clinical application of selected cell-based tumor vaccines achieved due to the generation and development of gene transfer technologies.     

乳腺癌双膦酸盐辅助治疗临床研究进展

乳腺癌是女性发病率和死亡率最高的恶性肿瘤,复发和远处转移仍是导致患者死亡的首位原因,而双膦酸盐作为一种骨质吸收抑制剂,能够抑制破骨细胞介导的骨质吸收,在多种实体肿瘤骨转移及多发性骨髓瘤等恶性疾病所致的骨相关事件治疗中起重要作用。近年来大量体外、体内实验表明双膦酸盐还具有抑制肿瘤细胞生长、粘附、播散和侵润,降低肿瘤细胞膜稳定性、促进肿瘤细胞凋亡等直接抗肿瘤作用以及抑制肿瘤血管生成、激活免疫细胞对肿瘤细胞的杀伤等间接抗肿瘤作用,基于这些基础研究结果已经开展了一系列针对双膦酸盐辅助治疗乳腺癌的,陆床试验研究,本文就近年相关临床试验研究进展做简要综述。     

Potential of chimeric antigen receptor (CAR)‐redirected immune cells in breast cancer therapies: Recent advances

Despite substantial developments in conventional treatments such as surgery, chemotherapy, radiotherapy, endocrine therapy, and molecular‐targeted therapy, breast cancer remains the leading cause of cancer mortality in women. Currently, chimeric antigen receptor (CAR)–redirected immune cell therapy has emerged as an innovative immunotherapeutic approach to ameliorate survival rates of breast cancer patients by eliciting cytotoxic activity against cognate tumour‐associated antigens expressing tumour cells. As a crucial component of adaptive immunity, T cells and NK cells, as the central innate immune cells, are two types of pivotal candidates for CAR engineering in treating solid malignancies. However, the biological distinctions between NK cells‐ and T cells lead to differences in cancer immunotherapy outcomes. Likewise, optimal breast cancer removal via CAR‐redirected immune cells requires detecting safe target antigens, improving CAR structure for ideal immune cell functions, promoting CAR‐redirected immune cells filtration to the tumour microenvironment (TME), and increasing the ability of these engineered cells to persist and retain within the immunosuppressive TME. This review provides a concise overview of breast cancer pathogenesis and its hostile TME. We focus on the CAR‐T and CAR‐NK cells and discuss their significant differences. Finally, we deliver a summary based on recent advancements in the therapeutic capability of CAR‐T and CAR‐NK cells in treating breast cancer.     

Immunologic biomarkers as correlates of clinical response to cancer immunotherapy

Over the last few years, several newly developed immune-based cancer therapies have been shown to induce clinical responses in significant numbers of patients. As a result, there is a need to identify immune biomarkers capable of predicting clinical response. If there were laboratory parameters that could define patients with improved disease outcomes after immunomodulation, product development would accelerate, optimization of existing immune-based treatments would be facilitated and patient selection for specific interventions might be optimized. Although there are no validated cancer immunologic biomarkers that are predictive of clinical response currently in widespread use, there is much published literature that has informed investigators as to which markers may be the most promising. Population-based studies of endogenous tumor immune infiltrates and gene expression analyses have identified specific cell populations and phenotypes of immune cells that are most likely to mediate anti-tumor immunity. Further, clinical trials of cancer vaccines and other cancer directed immunotherapy have identified candidate immunologic biomarkers that are statistically associated with beneficial clinical outcomes after immune-based cancer therapies. Biomarkers that measure the magnitude of the Type I immune response generated with immune therapy, epitope spreading, and autoimmunity are readily detected in the peripheral blood and, in clinical trials of cancer immunotherapy, have been associated with response to treatment.     

Murine model of hepatic breast cancer

Background and aimsBreast cancer is the most common cancer in women and the second leading cause of cancer-related deaths in this population. Breast cancer related deaths have declined due to screening and adjuvant therapies, yet a driving clinical need exists to better understand the cause of the deadliest aspect of breast cancer, metastatic disease. Breast cancer metastasizes to several distant organs, the liver being the third most common site. To date, very few murine models of hepatic breast cancer exist.MethodsIn this study, a novel murine model of liver breast cancer using the MDA-MB-231 cell line is introduced as an experimental (preclinical) model.ResultsHistological typing revealed consistent hepatic breast cancer tumor foci. Common features of the murine model were vascular invasion, lung metastasis and peritoneal seeding.ConclusionsThe novel murine model of hepatic breast cancer established in this study provides a tool to be used to investigate mechanisms of hepatic metastasis and to test potential therapeutic interventions.     

Ectodermal‐neural cortex 1 as a novel biomarker predicts poor prognosis and induces metastasis in breast cancer by promoting Wnt/β‐catenin pathway

Breast cancer, as the most common malignancy, is the second leading cause of cancer‐related death in women. One of the kelch family member ENC1 is involved in various pathophysiologic processes. But the role of ENC1 in breast cancer has not been investigated. The present study value the feature, clinical significance and the molecular mechanisms of ENC1 in breast cancer. The expression and prognosis value of ENC1 expression among breast cancer and normal breast tissue were investigated in The Cancer Genome Atlas database and human samples. ENC1 was knockdown to explore its function in various breast cancer cell lines. Western blot was performed to explore the potential molecular mechanisms. We observed that ENC1 was overexpressed in breast cancer tissues. ENC1 overexpression was associated with high metastasis and predicted a poor prognosis in patients with breast cancer. ENC1 Knockdown inhibits the growth, clone formation, migration and invasion of breast cancer cells. Mechanism analysis revealed ENC1 was strong associated with the metastasis by modulating β‐catenin pathway. Our study emphasizes that ENC1 is a potential prognostic and metastasis‐related marker of breast cancer, and may function as a possible therapeutic target against breast cancer.     

乳腺癌双膦酸盐辅助治疗临床研究进展

乳腺癌是女性发病率和死亡率最高的恶性肿瘤,复发和远处转移仍是导致患者死亡的首位原因,而双膦酸盐作为一种骨质吸收抑制剂,能够抑制破骨细胞介导的骨质吸收,在多种实体肿瘤骨转移及多发性骨髓瘤等恶性疾病所致的骨相关事件治疗中起重要作用。近年来大量体外、体内实验表明双膦酸盐还具有抑制肿瘤细胞生长、粘附、播散和侵润,降低肿瘤细胞膜稳定性、促进肿瘤细胞凋亡等直接抗肿瘤作用以及抑制肿瘤血管生成、激活免疫细胞对肿瘤细胞的杀伤等间接抗肿瘤作用,基于这些基础研究结果已经开展了一系列针对双膦酸盐辅助治疗乳腺癌的临床试验研究,本文就近年相关临床试验研究进展做简要综述。     

Plumbagin-induced apoptosis of human breast cancer cells is mediated by inactivation of NF-kappaB and Bcl-2

Breast cancer remains the major cause of cancer-related deaths in women world-wide. The heterogeneity of breast cancer has further complicated the progress of target-based therapies. Triple negative breast cancers, lacking estrogen receptor, progesterone receptor and the Her-2/neu (ErbB2), represent a highly aggressive breast cancer subtype, that are difficult to treat. Pleiotropic agents, such as those found in nature, can target receptor-positive as well as receptor-negative cancer cells, suggesting that such agents could have significant impact in breast cancer prevention and/or therapy. Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone) is one such agent which has anti-tumor activity against several cancers. However, its mechanism of action against breast cancer is not clearly understood. We hypothesized that plumbagin may act as an effective agent against breast cancer especially triple negative breast cancer. We tested our hypothesis using ER-positive MCF-7 and ER-negative MDA-MB-231 (triple negative) breast cancer cells, and we found that plumbagin significantly inhibits the growth of breast cancer cells with no effect on normal breast epithelial cells. We also found that plumbagin induces apoptosis with concomitant inactivation of Bcl-2 and the DNA binding activity of NF-kappaB. Bcl-2 over-expression resulted in attenuation of plumbagin-induced effects, suggesting that the inhibition of cell growth and induction of apoptosis by plumbagin is in part due to inactivation of NF-kappaB/Bcl-2 pathway. To our knowledge, this is the first report, showing mechanistic and cancer cell specific apoptosis-inducing effects of plumbagin in breast cancer cells, suggesting the potential role of plumbagin in the prevention and/or treatment of breast cancer.     

Multifaceted Roles of Interleukin-6 in Adipocyte–Breast Cancer Cell Interaction

Breast cancer is the most common malignancy in women worldwide, with a developmental process spanning decades. The malignant cells recruit a variety of cells including fibroblasts, endothelial cells, immune cells, and adipocytes, creating the tumor microenvironment. The tumor microenvironment has emerged as active participants in breast cancer progression and response to treatment through autocrine and paracrine interaction with the malignant cells. Adipose tissue is abundant in the breast cancer microenvironment; interactions with cancer cells create cancer-associated adipocytes which produce a variety of adipokines that influence breast cancer initiation, metastasis, angiogenesis, and cachexia. Interleukin (IL)-6 has emerged as key compound significantly produced by breast cancer cells and adipocytes, with the potential of inducing proliferation, epithelial-mesenchymal phenotype, stem cell phenotype, angiogenesis, cachexia, and therapeutic resistance in breast cancer cells. Our aim is to present a brief knowledge of IL-6’s role in breast cancer. This review summarizes our current understanding of the breast microenvironment, with emphasis on adipocytes as key players in breast cancer tumorigenesis. The effects of key adipocytes such as leptin, adipokines, TGF-b, and IL-6 are discussed. Finally, we discuss the role of IL-6 in various aspects of cancer progression.     

乳腺癌的治疗进展

乳腺癌是女性最常见的恶性肿瘤之一,是一种严重影响女性身心健康甚至危及生命的恶性肿瘤,男性乳腺癌罕见。目前,乳腺癌的治疗仍以外科治疗为主。乳腺癌的各种治疗手段,如手术治疗、化学治疗、放射治疗、内分泌治疗等,依据各自特点及疗效,在治疗中综合运用,使乳腺癌的临床疗效有了明显提高。随着治疗模式的改变、概念的不断更新以及越来越多的相关治疗机制被发现,能提高患者的综合治疗效果及生活质量,延长生存期。本文根据国内外已发表的乳腺癌治疗相关文献,主要从乳腺癌治疗相关进展及治疗效果等方面进行归纳总结,为制订乳腺癌综合治疗的指南提供一些参考。     

Tumor markers in breast cancer--evaluation of their clinical usefulness

Breast cancer is the most common neoplasm affecting women in the Western world. Many studies are still conducted with the purpose of finding markers that could be used for early diagnosis and/or serve as possible reliable prognostic or predictive parameters, but with conflicting results. At present, no markers are available for an early diagnosis of breast cancer For surveillance of patients with diagnosed breast cancer the most widely used serum markers are CA 15-3 and CEA which, in combination with other clinical parameters, could have clinical significance. The most useful and clinically important tissue-based markers in breast cancer are estrogen and progesterone receptors, used as a basis for hormonal therapy, and HER-2 receptors, essential in selecting patients for the treatment with Herceptin. New or potentially new markers for breast cancer include BRCA1 and BRCA2 genes for selecting patients at high risk of developing hereditary breast cancer, as well as urokinase plasminogen activator and inhibitor for assessing prognosis in lymph node-negative patients. Results of tumor and patient genetic analyses including their clinical evaluation will enable application of more individualized and personalized approach in diagnosis and therapy of breast cancer patients.