Light-controllable charge-reversal nanoparticles with polyinosinic-polycytidylic acid for enhancing immunotherapy of triple negative breast cancer

Nucleic acid drugs are highly applicable for cancer immunotherapy with promising therapeutic effects, while targeting delivery of these drugs to disease lesions remains challenging. Cationic polymeric nanoparticles have paved the way for efficient delivery of nucleic acid drugs, and achieved stimuli-responsive disassembly in tumor microenvironment (TME). However, TME is highly heterogeneous between individuals, and most nanocarriers lack active-control over the release of loaded nucleic acid drugs, which will definitely reduce the therapeutic efficacy. Herein, we have developed a light-controllable charge-reversal nanoparticle (LCCN) with controlled release of polyinosinic-polycytidylic acid [Poly(I:C)] to treat triple negative breast cancer (TNBC) by enhanced photodynamic immunotherapy. The nanoparticles keep suitably positive charge for stable loading of Poly(I:C), while rapidly reverse to negative charge after near-infrared light irradiation to release Poly(I:C). LCCN-Poly(I:C) nanoparticles trigger effective phototoxicity and immunogenic cell death on 4T1 tumor cells, elevate antitumor immune responses and inhibit the growth of primary and abscopal 4T1 tumors in mice. The approach provides a promising strategy for controlled release of various nucleic acid-based immune modulators, which may enhance the efficacy of photodynamic immunotherapy against TNBC     

Curcumin-derivative nanomicelles for the treatment of triple negative breast cancer

Abstract

Background: Triple negative breast cancer (TNBC) is a subtype of breast cancer characterized by its poor outcome and a lack of targeted therapies. Recently, our laboratory has developed a second generation curcumin derivative, 3,5-bis(3,4,5-trimethoxybenzylidene)-1-methylpiperidine-4-one (RL71) that shows potent in vitro cytotoxicity. RL71 is hydrophobic with poor bioavailability which limits its clinical development.

Purpose: We have designed styrene-co-maleic acid (SMA) micelles encapsulating 5, 10 or 15% RL71 by weight/weight ratio to improve its solubility and pharmacokinetic profile.

Methods: The micelles charge, size and release rate were characterized. We evaluated their cytotoxicity against TNBC cell lines. The internalization of the drug inside the cells was measured by HPLC and the efficiency of the micelles was tested using a tumor spheroid model.

Results: The micelles exhibited mean diameters of 125–185?nm and had a neutral charge. SMA-RL71 micelles have a cytotoxicity profile comparable to the free drug against several TNBC cell lines. Moreover, the 15% loaded micelles increased the stability of RL71 and demonstrated higher activity in a tumor spheroid model.

Conclusion: The current study demonstrates the efficiency of SMA for drug delivery, the influence of physicochemical characteristics on cytotoxicity, and provides the basis for preclinical testing in vivo.     

Discovery of a small molecule targeting ULK1-modulated cell death of triple negative breast cancer in vitro and in vivo

OBJECTIVE To discover a small molecule targeting ULK1-modulated cell death of triple negative breast cancer and exploreits potential mechanisms.METHODS ULK1 expression was analyzed by The Cancer Genome Atlas(TCGA)analysis and tissue microarray(TMA)analysis.ULK1agonist was designed by using in silico screening,as well as modified by chemical synthesis and screened by kinase and anti-proliferative activities.The amino acid residues that key to the activation site of LYN-1604 were determined by site-directed mutagenesis,as well as in vitro kinase assay and ADP-Glo kinase assay.The mechanisms of LYN-1604 induced cell death were investigated by fluorescence microscope,western blotting,flow cytometry analysis,immunocytochemistry,as well as si RNA and GFP-m RFP-LC3 plasmid transfections.Potential ULK1 interactors were discovered by performing comparative microarray analysis and the therapeutic effect of LYN-1604 was assessed by xenograft breast cancer mouse model.RESULTS We found that ULK1 was remarkably downregulated in breast cancer tissue samples,especial y in triple negative breast cancer(TNBC).32 candidate smal molecules were synthesized,and we discovered a small molecule named LYN-1604 as the best candidate ULK1agonist.Additionally,we identified that three amino acid residues(LYS50,LEU53 and TYR89)were key to the activation site of LYN-1604 and ULK1.Subsequently,we demonstrated that LYN-1604 could induce autophagy-associated cell death via ULK complex(ULK1-m ATG13-FIP200-ATG101)in MDA-MB-231 cells.We also found that LYN-1604 induced cell death involved in ATF3,RAD21 and caspase 3,accompanied with autophagy and apoptosis.Moreover,we demonstrated that LYN-1604 had a good therapeutic potential on TNBC by targeting ULK1-modulated cell death in vivo.CONCLUSION We discovered a small molecule(LYN-1604)has therapeutic potential by targeting ULK1-modulated cell death associated with autophagy and apoptosis of TNBC in vitro and in vivo,which could be utilized as a new anti-TNBC drug candidate.     

长春瑞滨联合顺铂治疗三阴性乳腺癌的临床研究

目的探讨长春瑞滨联合顺铂治疗三阴性乳腺癌的临床疗效和安全性。方法对我院2006年3月至2009年1月收治的41例明确诊断为三阴性乳腺癌的患者采用长春瑞滨联合顺铂治疗,评价患者化疗后的疗效和不良反应。结果临床疗效CR 7.3%(3/41)、PR 36.6%(15/41)、SD 36.6%(15/41)、PD 19.5%(8/41)、RR 43.9%(18/41)。化疗后随访至2012年2月,5例失访,中位TTP为6.0个月,不同转移部位的患者有效率不同,淋巴结和骨转移有效率最高,分别为60.9%(14/23)和38.9%(7/18)。患者化疗后不良反应主要为骨髓抑制、胃肠道反应、静脉炎等,未见化疗相关的死亡病例。结论长春瑞滨联合顺铂治疗三阴性乳腺癌临床疗效较好,且患者可耐受,值得临床进一步探讨和研究。     

Use of nanotechnology for improved pharmacokinetics and activity of immunogenic cell death inducers used in cancer chemotherapy

Introduction: Immunogenic cell death inducers (ICD inducers) are a diverse group of therapeutic molecules capable of eliciting an adaptive immune response against the antigens present on the surface of dying cancer cells. Most of these molecules suffer from low bioavailability, high toxicity and poor pharmacokinetics which limit their application. It is believed that nanotechnology, in particular nano-sized nanocarriers, can address most of the issues that limit the use of ICD inducers.

Area covered: The mechanism of action of ICD inducers and their limitations is discussed. In addition, we cover the novel possibilities arising from the use of nanotechnology to improve delivery of ICD inducers to the target tissue as well as the restrictions of modern nanotechnology.

Expert opinion: At present, nanocarrier formulations suffer from low bioavailability, poor pharmacokinetics and stability issues. Nonetheless, there is a tremendous future for combinatorial immune-pharmacological treatments of human tumors based on nanocarrier delivery of ICD inducers.     

The potential of Beclin 1 as a therapeutic target for the treatment of breast cancer

Introduction: Beclin 1 plays a crucial role in autophagy via the Beclin 1 interactome, and is involved in various biological processes such as protein sorting, chemokinesis, and cell death. Via these biologic functions, Beclin 1 contributes to both tumor suppression and tumor progression.

Areas covered: Beclin 1 plays a key biologic function on cell homeostasis and affects tumorigenesis. In this review, detailing up-to-date knowledge on the tumorigenic role of Beclin 1, its implication in breast cancer, and its utility as a breast cancer-specific drug target is discussed.

Expert opinion: Because Beclin 1 is expressed in breast cancer cells, Beclin 1 could be a unique, effective drug target for the prevention and treatment of breast cancer. However, the expression of Beclin 1 varies according to cancer molecular subtypes, and Beclin 1 is involved in both breast cancer suppression and tumor progression; therefore, the decision of using a Beclin 1 inducer or inhibitor should be made based on breast cancer stage and subtype.     

DNA/RNA-based formulations for treatment of breast cancer

Introduction: To develop a successful formulation for the gene therapy of breast cancer, an effective therapeutic nucleic acid and a proper delivery system are essential. Increased understanding of breast cancer, and developments in biotechnology, material science and nanotechnology have provided a major impetus in the development of effective formulations for the gene therapy of breast cancer.

Areas covered: We discuss DNA/RNA-based formulations that can inhibit the growth of breast cancer cells and control the progress of breast cancer. Targets for the gene therapy of breast cancer, DNA/RNA-based therapeutics and delivery systems are summarized. And examples of successful DNA/RNA-based formulations for breast cancer gene therapy are reviewed.

Expert opinion: Several challenges remain in developing effective DNA/RNA-based formulations for treatment of breast cancer. Firstly, most of the currently utilized targets are not effective enough as monotherapy for breast cancer. Secondly, the requirements for co-delivery system make the preparation of formulation more complicated. Thirdly, nanoparticles with the modification of tumor-targeting ligands could be more unstable in circulation and normal tissues. Lastly, immune responses against the viral vectors are unfavorable for the gene therapy of breast cancer because of the damage to the host and the impaired therapeutic ability.     

参附注射液联合表柔比星和紫杉醇治疗三阴性乳腺癌的临床研究

目的探究参附注射液联合表柔比星和紫杉醇治疗三阴性乳腺癌的临床疗效。方法选择2011年5月—2014年5月在海南省人民医院接受治疗的三阴性乳腺癌患者150例,随机分为对照组和治疗组,每组各75例。对照组在每个化疗周期的第1天静脉滴注注射用盐酸表柔比星60 mg/m2,1次/d,同时静脉滴注紫杉醇注射液120 mg/m2,1次/d,滴注时间大于3 h。治疗组在每个化疗周期的1~10 d静脉滴注参附注射液50 m L/次,注射用盐酸表柔比星和紫杉醇注射液的用法用量同对照组。21 d为一个化疗疗程,两组均连续治疗2个疗程。治疗后,评价两组的临床疗效,同时比较两组患者治疗前后肿瘤直径、癌细胞的叉头框蛋白A1(fork box protein A1,FOXA1)、乳腺癌易感基因(breast cancer susceptibility gene 1,BRCA1)蛋白水平的变化。结果治疗后,对照组和治疗组的总有效率分别为82.67%、64.00%,两组比较差异有统计学意义(P0.05)。治疗后,两组患者的肿瘤直径、FOXA1、BRCA1蛋白均较治疗前显著降低,同组治疗前后差异有统计学意义(P0.05);且治疗组的降低程度优于对照组,两组比较差异具有统计学意义(P0.05)。对照组和治疗组的不良反应发生率分别为18.67%、13.33%,两组比较差异有统计学意义(P0.05)。结论参附注射液联合表柔比星和紫杉醇治疗三阴性乳腺癌效果较好,可显著降低患者肿瘤直径,还可降低FOXAl和BRCAl蛋白水平,且不良反应较低,在临床具有一定的推广应用价值。     

Dual-responsive nanoparticles with transformable shape and reversible charge for amplified chemo-photodynamic therapy of breast cancer

Herein, we designed a dual-response shape transformation and charge reversal strategy with chemo-photodynamic therapy to improve the blood circulation time, tumor penetration and retention, which finally enhanced the anti-tumor effect. In the system, hydrophobic photosensitizer chlorin e6 (Ce6), hydrophilic chemotherapeutic drug berberrubine (BBR) and matrix metalloproteinase-2 (MMP-2) response peptide (PLGVRKLVFF) were coupled by linkers to form a linear triblock molecule BBR-PLGVRKLVFF-Ce6 (BPC), which can self-assemble into nanoparticles. Then, positively charged BPC and polyethylene glycol-histidine (PEG-His) were mixed to form PEG-His@BPC with negative surface charge and long blood circulation time. Due to the acidic tumor microenvironment, the PEG shell was detached from PEG-His@BPC attributing to protonation of the histidine, which achieved charge reversal, size reduction and enhanced tumor penetration. At the same time, enzyme cutting site was exposed, and the spherical nanoparticles could transform into nanofibers following the enzymolysis by MMP-2, while BBR was released to kill tumors by inducing apoptosis. Compared with original nanoparticles, the nanofibers with photosensitizer Ce6 retained within tumor site for a longer time. Collectively, we provided a good example to fully use the intrinsic properties of different drugs and linkers to construct tumor microenvironment-responsive charge reversal and shape transformable nanoparticles with synergistic antitumor effect.     

注射用香菇多糖联合AC方案和紫杉醇治疗晚期三阴性乳腺癌的临床研究

目的评价注射用香菇多糖联合AC方案和紫杉醇治疗晚期三阴性乳腺癌的临床疗效和安全性。方法选取2015年1月1日—2017年12月1日天津中医药大学第一附属医院和天津医科大学附属肿瘤医院收治的60例乳腺癌患者为研究对象,将所有患者随机分为对照组和治疗组,每组各30例。对照组患者采用单纯密集AC方案联合紫杉醇:第1天静脉滴注注射用盐酸多柔比星,60 mg/m~2,同时第1天也静脉推注注射用环磷酰胺,600 mg/m~2,14 d为1个周期,治疗4个周期;之后序贯第1天静脉滴注注射用紫杉醇(白蛋白结合型),175 mg/m~2,14 d为1个周期,治疗4个周期。治疗组在对照组治疗的基础上静脉注射注射用香菇多糖,1 mg用2 mL注射用水振摇溶解,加入到生理盐水250 mL中,2次/周,14 d为1个周期,治疗4个周期后进行观察。观察两组的近期临床疗效,比较两组的生活质量评分、淋巴细胞亚群水平、不良反应情况。结果治疗后,对照组和治疗组的客观有效率(ORR)分别为56.7%、63.3%,疾病控制率(DCR)分别为86.7%、90.0%,两组比较差异无有统计学意义。治疗后,两组患者KPS评分升高,同组治疗前后比较差异无统计学意义,且两组治疗后比较差异亦无统计学意义。治疗后,对照组患者CD4~+/CD8~+水平显著降低,治疗组患者CD4~+、CD4~+/CD8~+水平均显著升高,同组治疗前后比较差异具有统计学意义(P0.05);且治疗后治疗组CD4~+、CD4~+/CD8~+水平明显高于对照组,两组治疗后比较差异具有统计学意义(P0.05)。治疗后,治疗组白细胞降低、恶心呕吐的Ⅲ～Ⅳ发生率均明显低于对照组,两组不良反应发生率比较差异有统计学意义(P0.05)。结论注射用香菇多糖联合AC方案和紫杉醇治疗晚期三阴性乳腺癌具有较好的临床疗效,能改善患者淋巴细胞亚群水平,降低不良反应,具有一定的临床推广应用价值。     

Co-delivery of gambogic acid and TRAIL plasmid by hyaluronic acid grafted PEI-PLGA nanoparticles for the treatment of triple negative breast cancer

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-based combination therapy and gene therapy are new strategies to potentially overcome the limitations of TRAIL, however, the lack of efficient and low toxic vectors remains the major obstacle. In this study, we developed a hyaluronic acid (HA)-decorated polyethylenimine-poly(d,l-lactide-co-glycolide) (PEI-PLGA) nanoparticle (NP) system for targeted co-delivery of TRAIL plasmid (pTRAIL) and gambogic acid (GA) in triple-negative breast cancer (TNBC) therapy. GA was encapsulated into the core of the PEI-PLGA NPs while pTRAIL was adsorbed onto the positive NP surface via charge adsorption. The coating of HA on PEI-PLGA NPs functions as a targeting ligand by binding to CD44 receptor of TNBC cells and a shell to neutralize the excess positive charge of inner NPs. The resultant pTRAIL and GA co-loaded HA-coated PEI-PLGA NPs exhibited spherical shape (121.5?nm) and could promote the internalization of loaded cargoes into TNBC cells through the CD44-dependent endocytic pathway. The dual drug-loaded NPs significantly augmented apoptotic cell death in vitro and inhibited TNBC tumor growth in vivo. This multifunctional NP system efficiently co-delivered GA and pTRAIL, thus representing a promising strategy to treat TNBC and bringing forth a platform strategy for co-delivery of therapeutic DNA and chemotherapeutic agents in combinatorial TNBC therapy.     

Nanoparticles for imaging and treatment of metastatic breast cancer

Introduction: Metastatic breast cancer is one of the most devastating cancers that have no cure. Many therapeutic and diagnostic strategies have been extensively studied in the past decade. Among these strategies, cancer nanotechnology has emerged as a promising strategy in preclinical studies by enabling early identification of primary tumors and metastases, and by effective killing of cancer cells.

Areas covered: This review covers the recent progress made in targeting and imaging of metastatic breast cancer with nanoparticles, and treatment using nanoparticle-enabled chemo-, gene, photothermal- and radio-therapies. This review also discusses recent developments of nanoparticle-enabled stem cell therapy and immunotherapy.

Expert opinion: Nanotechnology is expected to play important roles in modern therapy for cancers, including metastatic breast cancer. Nanoparticles are able to target and visualize metastasis in various organs, and deliver therapeutic agents. Through targeting cancer stem cells, nanoparticles are able to treat resistant tumors with minimal toxicity to healthy tissues/organs. Nanoparticles are also able to activate immune cells to eliminate tumors. Owing to their multifunctional, controllable and trackable features, nanotechnology-based imaging and therapy could be a highly potent approach for future cancer research and treatment.     

国产多西紫杉醇联合顺铂在晚期非小细胞肺癌中的应用

目的观察国产多西紫杉醇联合顺铂治疗晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的近期疗效和不良反应。方法采用DP方案化疗:多帕菲75mg/m2,静脉滴注,第1天;顺铂75mg/m2,静脉滴注,第1天;21d为1个周期。至少使用2个周期评价疗效。结果70例中CR2例,PR29例,SD36例,PD3例,总有效率为44.3%。中位TTP6.3个月,中位生存期10.1个月,1年生存率38.6%。不良反应主要为骨髓抑制、脱发和消化系统反应,骨髓抑制为剂量限制性毒性,其中白细胞减少占75.7%。结论国产多西紫杉醇联合顺铂治疗晚期NSCLC近期疗效较好,毒不良反应较轻,值得临床进一步推广应用。     

干预雌激素代谢防治乳腺癌的药物研究进展

乳腺癌是女性排名第一的常见恶性肿瘤,是一种激素相关性疾病,暴露于雌激素是患乳腺癌一个重要的决定因素。而相关研究表明,干预雌激素的代谢过程,能够达到有效防治乳腺癌的目的。在雌激素代谢过程中,2-羟基雌激素(2-OHE)、2-甲氧基雌激素(2-OMeE)、4-甲氧基雌激素(4-OMeE)为有利代谢产物,16-羟基雌激素(16-OHE)、4-羟基雌激素(4-OHE)为有害代谢产物。若提高2-OHE与16-OHE含量比,可有效防治乳腺癌。此外,提高儿茶酚邻位甲基转移酶活性用以防治乳腺癌也具有广阔的前景。就近年来对于雌激素代谢过程和基于雌激素代谢防治乳腺癌的研究进展进行综述。     

多西他赛联合顺铂治疗晚期非小细胞肺癌43例

目的:观察国产多西他赛联合顺铂治疗晚期非小细胞肺癌的疗效及不良反应。方法:不能手术的Ⅲ、Ⅳ期非小细胞肺癌病人43例。d 1多西他赛75 mg·m~(-2),顺铂70～75 mg·m~(-2),21 d为一个周期,每2个周期评价疗效,共完成4个周期。结果:43例病人共完成165个周期化疗,均可评价疗效,完全缓解(CR)1例,部分缓解(PR)13例,微效(MR)7倒,疾病稳定(SD)13例,疾病进展(PD)9例,总有效率(RR)49％,临床获益率79％,中位生存期(MST)是10．7 mo,疾病进展时间(TTP)是7．2 mo,1年生存率是39％。主要不良反应是白细胞下降、消化道反应、乏力、脱发。结论:多西他赛联合顺铂治疗晚期非小细胞肺癌疗效确切,耐受良好。     

Coordinated regulation of BACH1 and mitochondrial metabolism through tumor-targeted self-assembled nanoparticles for effective triple negative breast cancer combination therapy

The poor prognosis of triple negative breast cancer (TNBC) results from a lack of approved targeted therapies coupled with aggressive proliferation and metastasis, which is associated with high recurrence and short overall survival. Here we developed a strategy by employing tumor-targeted self-assembled nanoparticles to coordinately regulate BACH1 (BTB domain and CNC homology 1) and mitochondrial metabolism. The BACH1 inhibitor hemin and mitochondria function inhibitor berberine derivative (BD) were used to prepare nanoparticles (BH NPs) followed by the modification of chondroitin sulfate (CS) on the surface of BH NPs to achieve tumor targeting (CS/BH NPs). CS/BH NPs were found to be able to inhibit tumor migration and invasion by significantly decreasing the amounts of tumor cell metabolites, glycolysis and metastasis-associated proteins, which were related to the inhibition of BACH1 function. Meanwhile, decreased mitochondrial membrane potential, activated caspase 3/9 and increased ROS production demonstrated coordinated regulation of BACH1 and mitochondrial metabolism. In a xenograft mice model of breast cancer, CS/BH NPs significantly inhibited tumor growth and metastasis due to the synergetic effect of hemin and BD without showing obvious toxicities for major organs. In sum, the results of efficacy and safety experiments suggest potential clinical significance of the prepared self-assembled CS/BH nanoparticles for the treatment of TNBC.     

CD44 directed nanomicellar payload delivery platform for selective anticancer effect and tumor specific imaging of triple negative breast cancer

Triple negative breast cancer (TNBC) is a highly aggressive tumor subtype, lacking estrogen, progesterone and human epidermal growth factor-2 (HER-2) receptors. Thus, early detection and targeted therapy of TNBC is an urgent need. Herein, we have developed a CD44 targeting Hyaluronic Acid (HA) decorated biocompatible oligomer, containing FDA approved vitamin E TPGS and Styrene Maleic Anhydride (SMA) (HA-SMA-TPGS) for targeting TNBC. The self-assembling HA-SMA-TPGS was encapsulated with poorly water soluble, potent curcumin analogue (CDF) to form nanomicelles (NM), HA-SMA-TPGS-CDF has demonstrated excellent nanoparticle characteristics for parenteral delivery. The targeted NM can selectively kill TNBC cells through CD44 mediated apoptosis pathway. Tumor imaging using phase-2 clinical trial near infrared (NIR)-fluorescent dye (S0456) conjugate, HA-SMA-TPGS-S0456 showed excellent TNBC tumor accumulation with minimum liver and spleen uptake. To our best of knowledge, for the first time, we are reporting a promising platform for CD44 mediated multimodal NIR imaging and cytotoxin delivery to TNBC.     

The cancer stem cell paradigm: a new understanding of tumor development and treatment

Importance of the field: Cancer is the second leading cause of death in the United States, and therefore remains a central focus of modern medical research. Accumulating evidence supports a ‘cancer stem cell’ (CSC) model – where cancer growth and/or recurrence is driven by a small subset of tumor cells that exhibit properties similar to stem cells. This model may provide a conceptual framework for developing more effective cancer therapies that target cells propelling cancer growth.

Areas covered in this review: We review evidence supporting the CSC model and associated implications for understanding cancer biology and developing novel therapeutic strategies. Current controversies and unanswered questions of the CSC model are also discussed.

What the reader will gain: This review aims to describe how the CSC model is key to developing novel treatments and discusses associated shortcomings and unanswered questions.

Take home message: A fresh look at cancer biology and treatment is needed for many incurable cancers to improve clinical prognosis for patients. The CSC model posits a hierarchy in cancer where only a subset of cells drive malignancy, and if features of this model are correct, has implications for development of novel and hopefully more successful approaches to cancer therapy.     

Lenvatinib-and vadimezan-loaded synthetic high-density lipoprotein for combinational immunochemotherapy of metastatic triple-negative breast cancerOA

Metastatic triple-negative breast cancer(TNBC) is the most aggressive type of breast cancer.Combination of systemic chemotherapy and immune checkpoint blockade is effective but of limited benefit due to insufficient intratumoral infiltration of cytotoxic T lymphocytes(CTLs) and the accumulation of immunosuppressive cells.Herein,we designed a lenvatinib-and vadimezan-loaded synthetic high-density lipoprotein(LV-sHDL) for combinational immunochemotherapy of metastatic TNBC.The LV-sHDL targeted sca...