Jendryczko Karolina, Rzeszotko Jakub, Krzyscik Mateusz Adam, Szymczyk Jakub, Otlewski Jacek, Szlachcic Anna
Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland.
Front Pharmacol. 2021 Nov 12;12:748936. doi: 10.3389/fphar.2021.748936. eCollection 2021.
Targeted therapies are a promising alternative to conventional chemotherapy, with an increasing number of therapeutics targeting specific molecular aberrancies in cancer cells. One of the emerging targets for directed cancer treatments is fibroblast growth factor receptors (FGFRs), which are known to be involved in the pathogenesis and progression of multiple cancer types, specially in lung, bladder, and breast cancers. Here, we are demonstrating the development of the FGFR1-targeting agent based on the interactome screening approach, based on the isolation of binding regions from ligands interacting with the receptor. The parallel analysis by FGFR1 pull-down of chymotryptic peptides coupled with MS analysis, and PepSpot analysis yielded equivalent peptide sequences from FGF4, one of the FGFR1 ligands. Three sequences served as a basis for peptibody (Fc-fusion) generation, to overcome clinical limitations of peptidic agents, and two of them showed favorable FGFR1-binding and FGFR1-dependent internalization into cells. To validate if developed FGFR1-targeting peptibodies can be used for drug delivery, similar to the well-established concept of antibody-drug conjugates (ADCs), peptibodyF4_1 was successfully conjugated with monomethylauristatin E (MMAE), and has shown significant and specific toxicity toward FGFR1-expressing lung cancer cell lines, with nanomolar EC values. Essentially, the development of new effective FGFR1 binders that comprise the naturally occurring FGFR-recognition peptides and Fc region ensuring high plasma stability, and long bloodstream circulation is an interesting strategy expanding targeted anticancer agents' portfolio. Furthermore, identifying peptides effectively binding the receptor from sequences of its ligands is not limited to FGFRs and is an approach versatile enough to be a basis for a new peptide/peptibodies development strategy.
靶向治疗是传统化疗的一种有前景的替代方法，越来越多的治疗药物针对癌细胞中的特定分子异常。定向癌症治疗的新兴靶点之一是成纤维细胞生长因子受体（FGFRs），已知其参与多种癌症类型的发病机制和进展，特别是在肺癌、膀胱癌和乳腺癌中。在此，我们展示了基于相互作用组筛选方法开发的FGFR1靶向剂，该方法基于从与受体相互作用的配体中分离结合区域。通过FGFR1下拉胰凝乳蛋白酶肽段并结合质谱分析和PepSpot分析进行的平行分析，从FGFR1配体之一FGF4中获得了等效的肽序列。三个序列作为生成肽抗体（Fc融合）的基础，以克服肽类药物的临床局限性，其中两个显示出良好的FGFR1结合以及FGFR1依赖性细胞内化。为了验证开发的FGFR1靶向肽抗体是否可用于药物递送，类似于成熟的抗体药物偶联物（ADC）概念，肽抗体F4_1成功与单甲基澳瑞他汀E（MMAE）偶联，并对表达FGFR1的肺癌细胞系显示出显著且特异性的毒性，其EC值为纳摩尔级。从本质上讲，开发新的有效FGFR1结合剂，其包含天然存在的FGFR识别肽和确保高血浆稳定性及长血液循环时间的Fc区域，是一种扩展靶向抗癌药物组合的有趣策略。此外，从其配体序列中鉴定有效结合受体的肽并不局限于FGFRs，并且是一种足够通用的方法，可作为新的肽/肽抗体开发策略的基础。
