Evaluation of variable new antigen receptors (vNARs) as a novel cathepsin S (CTSS) targeting strategy.
Evaluation of variable new antigen receptors (vNARs) as a novel cathepsin S (CTSS) targeting strategy.
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DOI：
10.3389/fphar.2023.1296567
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发表时间：
2023
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
--
作者：
半胱氨酸蛋白酶组织蛋白酶S(CTSS)的异常活性涉及广泛的病理过程。尤其在癌症中，CTSS被证明可以促进肿瘤的进展，主要是通过促进肿瘤细胞的侵袭和迁移以及增强血管生成。虽然是一个有吸引力的治疗靶点，但需要更有效的CTSS抑制剂。在这里，我们研究了可变的新抗原受体(VNAR)作为一种新的抑制策略的潜在应用。在针对人重组ProCTSS的噬菌体展示淘洗过程中，鉴定了一组潜在的vNAR结合子。随后对它们进行了表达、纯化和结合亲和力的酶联免疫吸附试验和SPR鉴定。将所选的铅克隆向前推进，并在重组酶活性检测中显示抑制CTSS活性。进一步的评估表明，我们的Lead克隆通过一种新的抑制机制发挥作用，阻止ProCTSS对成熟酶的激活。此外，使用体内方法，我们展示了这些克隆在细胞内表达并抑制CTSS活性的能力，同时在肿瘤细胞侵袭实验中也注意到铅克隆可以阻止细胞侵袭。总之，这些发现说明了一种抑制CTSS活性的新机制，抗CTSS vNAR克隆在对抗有害的CTSS活性方面具有治疗潜力。此外，这项研究例证了VNAR在靶向细胞内蛋白方面的潜力，为基于生物的治疗打开了一系列以前“无法下药”的靶点。
Aberrant activity of the cysteine protease Cathepsin S (CTSS) has been implicated across a wide range of pathologies. Notably in cancer, CTSS has been shown to promote tumour progression, primarily through facilitating invasion and migration of tumour cells and augmenting angiogenesis. Whilst an attractive therapeutic target, more efficacious CTSS inhibitors are required. Here, we investigated the potential application of Variable New Antigen Receptors (vNARs) as a novel inhibitory strategy. A panel of potential vNAR binders were identified following a phage display panning process against human recombinant proCTSS. These were subsequently expressed, purified and binding affinity confirmed by ELISA and SPR based approaches. Selected lead clones were taken forward and were shown to inhibit CTSS activity in recombinant enzyme activity assays. Further assessment demonstrated that our lead clones functioned by a novel inhibitory mechanism, by preventing the activation of proCTSS to the mature enzyme. Moreover, using an intrabody approach, we exhibited the ability to express these clones intracellularly and inhibit CTSS activity whilst lead clones were also noted to impede cell invasion in a tumour cell invasion assay. Collectively, these findings illustrate a novel mechanistic approach for inhibiting CTSS activity, with anti-CTSS vNAR clones possessing therapeutic potential in combating deleterious CTSS activity. Furthermore, this study exemplifies the potential of vNARs in targeting intracellular proteins, opening a range of previously “undruggable” targets for biologic-based therapy.
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影响因子：
7.3
作者：
Hilpert, Hans;Mauser, Harald;Haap, Wolfgang
通讯作者：
Haap, Wolfgang
影响因子：
4.7
作者：
Griffiths, Katherine;Dolezal, Olan;Foley, Michael
通讯作者：
Foley, Michael
影响因子：
6
作者：
Flannery, T;Gibson, D;Johnston, PG
通讯作者：
Johnston, PG
影响因子：
--
作者：
English, Hejiao;Hong, Jessica;Ho, Mitchell
通讯作者：
Ho, Mitchell
影响因子：
51.1
作者：
Entschladen, F;Drell, TL;Zaenker, KS
通讯作者：
Zaenker, KS