A novel humanized Frizzled-7-targeting antibody enhances antitumor effects of Bevacizumab against triple-negative breast cancer via blocking Wnt/β-catenin signaling pathway.
A novel humanized Frizzled-7-targeting antibody enhances antitumor effects of Bevacizumab against triple-negative breast cancer via blocking Wnt/β-catenin signaling pathway.
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一种新型人源化 Frizzled-7 靶向抗体通过阻断 Wnt/β-catenin 信号通路增强 Bevacizumab 对三阴性乳腺癌的抗肿瘤作用。
DOI：
10.1186/s13046-020-01800-x
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发表时间：
2021-01-12
期刊：
影响因子：
--
通讯作者：
Huang G
中科院分区：
文献类型：
--
作者：
Xie W;Zhao H;Wang F;Wang Y;He Y;Wang T;Zhang K;Yang H;Zhou Z;Shi H;Wang J;Huang G
抗血管生成治疗已广泛应用于恶性肿瘤的临床治疗。然而，这种治疗的有效性受到质疑，特别是在三阴性乳腺癌（TNBC）中。Bevacizumab是FDA批准的第一种抗血管生成药物，实际上增加了TNBC细胞的侵袭和转移特性，这是由于Wnt/β-catenin信号转导响应于缺氧而激活。作为Wnt/β-catenin信号传导的关键受体，Frizzled-7（Fzd 7）在TNBC中异常表达，表明Fzd 7是开发与抗血管生成剂组合的药物的潜在靶标。利用杂交瘤技术和抗体人源化技术制备Fzd 7靶向抗体（SHH 002-hu 1）。采用生物膜干涉法（BLI）和近红外（NIR）成像检测SHH 002-hu 1的亲和力和靶向性。接下来，我们验证了SHH 002-hu 1是否能够抑制贝伐单抗诱导的TNBC细胞的侵袭和迁移，并通过荧光素酶报告基因和western blot分析阐明了SHH 002-hu 1的分子机制。建立TNBC裸鼠移植瘤模型，以评估SHH 002-hu 1与贝伐单抗联合时的抗TNBC活性。然后，通过免疫荧光（IF）评估对推定的TNBC干细胞样细胞和Wnt/β-连环蛋白信号传导的影响。进一步通过裸鼠移植瘤模型和成球实验研究TNBC细胞的成瘤能力和自我更新能力。此外，通过检测血管生成拟态（VM）和缺氧诱导因子-1 α（HIF-1α）转录活性，评价SHH 002-hu 1对TNBC细胞缺氧适应的影响。新型人源化抗Fzd 7抗体（SHH 002-hu 1）与Fzd 7具有极高的亲和力，并特异性靶向Fzd 7+细胞和肿瘤组织。SHH 002-hu 1通过减弱Wnt/β-catenin信号通路抑制Bevacizumab诱导的TNBC细胞的侵袭、迁移和上皮-间充质细胞转化（EMT）。SHH 002-hu 1通过减少推定的TNBC干细胞样细胞的亚群，显著增强贝伐珠单抗抑制TNBC生长的能力，进一步减弱贝伐珠单抗增强的TNBC细胞的肿瘤起始和自我更新能力。SHH 002-hu 1通过阻断Wnt/β-catenin信号通路，有效抑制TNBC细胞对缺氧的适应。SHH 002-hu 1显著增强贝伐单抗的抗TNBC能力，并显示出预防TNBC复发的潜力，表明SHH 002-hu 1是与贝伐单抗协同治疗的良好候选者。在线版本包含补充材料，可通过10.1186/s13046-020-01800-x获得。
Anti-angiogenic therapy has been widely applied to the clinical treatment of malignant tumors. However, the efficacy of such treatments has been called into question, especially in triple-negative breast cancer (TNBC). Bevacizumab, the first anti-angiogenic agent approved by FDA, actually increases invasive and metastatic properties of TNBC cells, resulting from the activation of Wnt/β-catenin signaling in response to hypoxia. As a critical receptor of Wnt/β-catenin signaling, Frizzled-7 (Fzd7) is aberrantly expressed in TNBC, indicating Fzd7 a potential target for developing drugs to be combined with anti-angiogenic agents. Hybridoma technique and antibody humanization technique were utilized to generate a Fzd7-targeting antibody (SHH002-hu1). Biolayer interferometry (BLI) assay and near infrared (NIR) imaging were conducted to detect the affinity and targeting ability of SHH002-hu1. Next, whether SHH002-hu1 could suppress the invasion and migration of TNBC cells induced by Bevacizumab were validated, and the underlying molecular mechanisms were elucidated by luciferase reporter and western blot assays. The nude-mice transplanted TNBC models were established to assess the anti-TNBC activities of SHH002-hu1 when combined with Bevacizumab. Then, the effects on putative TNBC stem-like cells and Wnt/β-catenin signaling were evaluated by immunofluorescence (IF). Further, the tumor-initiating and self-renew capacity of TNBC cells were studied by secondary nude mouse xenograft model and sphere formation assay. In addition, the effects of SHH002-hu1 on the adaptation of TNBC cells to hypoxia were evaluated by the detection of vasculogenic mimicry (VM) and hypoxia-inducible factor-1α (HIF-1α) transcriptional activity. The novel humanized antibody targeting Fzd7 (SHH002-hu1) exhibited extremely high affinity with Fzd7, and specifically targeted to Fzd7+ cells and tumor tissues. SHH002-hu1 repressed invasion, migration and epithelial-mesenchymal cell transformation (EMT) of TNBC cells induced by Bevacizumab through abating Wnt/β-catenin signaling. SHH002-hu1 significantly enhanced the capacity of Bevacizumab to inhibit the growth of TNBC via reducing the subpopulation of putative TNBC stem-like cells, further attenuating Bevacizumab-enhanced tumor-initiating and self-renew capacity of TNBC cells. Moreover, SHH002-hu1 effectively restrained the adaptation of TNBC cells to hypoxia via disrupting Wnt/β-catenin signaling. SHH002-hu1 significantly enhances the anti-TNBC capacity of Bevacizumab, and shows the potential of preventing TNBC recurrence, suggesting SHH002-hu1 a good candidate for the synergistic therapy together with Bevacizumab. The online version contains supplementary material available at 10.1186/s13046-020-01800-x.
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影响因子：
11.1
作者：
McIntyre A;Harris AL
通讯作者：
Harris AL
影响因子：
11.2
作者：
Charafe-Jauffret E;Ginestier C;Iovino F;Wicinski J;Cervera N;Finetti P;Hur MH;Diebel ME;Monville F;Dutcher J;Brown M;Viens P;Xerri L;Bertucci F;Stassi G;Dontu G;Birnbaum D;Wicha MS
通讯作者：
Wicha MS
影响因子：
10.3
作者：
Lal, A;Peters, H;Riggins, GJ
通讯作者：
Riggins, GJ
DOI：
10.1056/nejmoa1111097
发表时间：
2012-01-26
期刊：
The New England journal of medicine
影响因子：
--
作者：
Bear HD;Tang G;Rastogi P;Geyer CE Jr;Robidoux A;Atkins JN;Baez-Diaz L;Brufsky AM;Mehta RS;Fehrenbacher L;Young JA;Senecal FM;Gaur R;Margolese RG;Adams PT;Gross HM;Costantino JP;Swain SM;Mamounas EP;Wolmark N
通讯作者：
Wolmark N
DOI：
10.1093/annonc/mdw544
发表时间：
2017-01-01
期刊：
Annals of oncology : official journal of the European Society for Medical Oncology
影响因子：
--
作者：
Cardoso F;Costa A;Senkus E;Aapro M;André F;Barrios CH;Bergh J;Bhattacharyya G;Biganzoli L;Cardoso MJ;Carey L;Corneliussen-James D;Curigliano G;Dieras V;El Saghir N;Eniu A;Fallowfield L;Fenech D;Francis P;Gelmon K;Gennari A;Harbeck N;Hudis C;Kaufman B;Krop I;Mayer M;Meijer H;Mertz S;Ohno S;Pagani O;Papadopoulos E;Peccatori F;Penault-Llorca F;Piccart MJ;Pierga JY;Rugo H;Shockney L;Sledge G;Swain S;Thomssen C;Tutt A;Vorobiof D;Xu B;Norton L;Winer E
通讯作者：
Winer E