The Cloning, Optimization, and Validation of Caninized-Phage Monoclonal Antibody Library For Regulation of Cancer Responses in Canus lupus

用于调节狼疮癌症反应的犬化噬菌体单克隆抗体库的克隆、优化和验证

• 批准号: BB/J00751X/1
• 负责人: Ted Hupp
• 金额: $ 52.07万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ00751X%2F1
• 关键词: Cloning; Optimization; Validation; Caninized; Phage

Cancer has emerged as a major cause of morbidity and mortality in the dog, with an estimated incidence of 1 in 4. Despite advances in conventional chemotherapy and radiation, complete cures for most cancer types remains elusive, the challenge being to develop highly targeted therapies that kill cancer cells but leave normal cells healthy. The same seemingly insurmountable problem holds true for the treatment of human cancer and a big advance in oncology in the past 20 years has been the discovery and application of monoclonal antibodies as novel therapeutic tools to improve cancer treatments in patients. In fact, one of the biggest areas of R and D in the pharmaceutical industry is now in the development of monoclonal antibodies. What are monoclonal antibodies (MAb's)? These are proteins made by our bodies that form a normal part of a healthy immune system and is the way nature helps to rid our bodies of foreign viruses, pathogenic bacteria, and also minimize disease development. Indeed, one reason why disease develops when we age or why we are more sensitive to viruses when we are older is essentially due to a "broken" immune system. However, scientists have discovered spectacular ways to harness the power of the immune system, by first cloning MAb's and then turning the MAb's on to treat particular diseases. The MAb's then act essentially like "magic bullets" as they can specifically hone in on rogue cells and cure disease. However, a major obstacle in this strategy of using MABs as magic bullets is that the immune system can recognize as foreign any particle that looks in any way different from normal proteins in the body; even a MAb given to patients with disease can paradoxically look foreign and be eliminated from the body before the MAb has a chance to act and elicit a cure. To overcome this obstacle, scientists in turn have pioneered a way to "tailor" the MAb so that it appears invisible to the body's immune system and can circulate and target disease cells. This tailoring of a MAb to treat humans is called "human-izing" the MAb and this approach provides further hope that complex diseases can be treated in the future. Despite these advances, the treatment of dog cancer, though in principle could be carried out like that in humans, has not been attempted in any significant way using MAb's. The main reason is that "dog-ized" MAb's would have to be used that are not recognized as invaders by the dog's immune system and dog-ized MAb's have not been developed that would greatly facilitate this process. In our research programme, we have developed novel methods to capture and clone the large majority of MAb's that can be made by the dog's immune system. Our application aims to optimize our dog-ized MAb library and validate it by isolating potential dog-ized "magic bullets" that can be used to inhibit specific receptor proteins that are implicated in causing dog cancers. The successful validation of these dog-ized MAb's will lead to clinical trials that would hopefully improve the health and longevity of dogs with cancer, but also produce a road map on how to develop dog-ized MAb's to treat other age related diseases in the dog.

癌症已成为狗发病和死亡的主要原因，估计发病率为四分之一。尽管传统化疗和放疗取得了进步，但大多数癌症类型的彻底治愈仍然难以实现，挑战在于开发高度靶向的疗法，杀死癌细胞但保持正常细胞健康。同样看似无法克服的问题也适用于人类癌症的治疗，过去 20 年肿瘤学的一大进步是单克隆抗体的发现和应用，作为新型治疗工具来改善患者的癌症治疗。事实上，目前制药行业最大的研发领域之一是单克隆抗体的开发。什么是单克隆抗体 (MAb)？这些是我们身体产生的蛋白质，构成健康免疫系统的正常部分，也是大自然帮助我们的身体清除外来病毒、致病细菌并最大程度减少疾病发展的方式。事实上，随着年龄的增长，我们会患上疾病，或者随着年龄的增长，我们对病毒更加敏感，原因之一本质上是由于免疫系统“受损”。然而，科学家们已经发现了利用免疫系统力量的惊人方法，首先克隆单克隆抗体，然后启动单克隆抗体来治疗特定疾病。单克隆抗体的作用本质上就像“神奇的子弹”，因为它们可以专门针对流氓细胞并治愈疾病。然而，这种使用 MAB 作为灵丹妙药的策略的一个主要障碍是，免疫系统可以将任何看起来与体内正常蛋白质有任何不同的颗粒识别为外来颗粒；即使是给予疾病患者的单克隆抗体也可能看起来很奇怪，并且在单克隆抗体有机会发挥作用并引起治愈之前就被从体内清除。为了克服这一障碍，科学家们又开创了一种“定制”单克隆抗体的方法，使其对人体的免疫系统来说是不可见的，并且可以循环并靶向疾病细胞。这种针对人类治疗的单克隆抗体被称为“人性化”单克隆抗体，这种方法为未来治疗复杂疾病提供了进一步的希望。尽管取得了这些进展，但狗癌症的治疗虽然原则上可以像人类一样进行，但尚未尝试使用单克隆抗体以任何有意义的方式进行治疗。主要原因是必须使用不被狗的免疫系统识别为入侵者的“狗化”单克隆抗体，并且尚未开发出能够极大促进这一过程的狗化单克隆抗体。在我们的研究项目中，我们开发了新的方法来捕获和克隆狗的免疫系统可以产生的绝大多数单克隆抗体。我们的应用旨在优化我们的狗化单克隆抗体库，并通过分离潜在的狗化“神奇子弹”来验证它，该“神奇子弹”可用于抑制与引起狗癌症有关的特定受体蛋白。这些狗源化单克隆抗体的成功验证将带来临床试验，有望改善患有癌症的狗的健康和寿命，同时也为如何开发狗源单克隆抗体来治疗狗的其他年龄相关疾病提供路线图。

项目成果

Reflux of Endoplasmic Reticulum proteins to the cytosol yields inactivation of tumor suppressors

内质网蛋白回流至细胞质导致肿瘤抑制因子失活

• DOI: 10.1101/2020.04.13.038935
• 发表时间: 2020
• 作者: Sicari D

Reflux of Endoplasmic Reticulum proteins to the cytosol inactivates tumor suppressors.

• DOI: 10.15252/embr.202051412
• 发表时间: 2021-05-05
• 期刊: EMBO reports
• 影响因子: 7.7
• 作者: Sicari D;Centonze FG;Pineau R;Le Reste PJ;Negroni L;Chat S;Mohtar MA;Thomas D;Gillet R;Hupp T;Chevet E;Igbaria A
• 通讯作者: Igbaria A

Hydrogen deuterium exchange mass spectrometry identifies the dominant paratope in CD20 antigen binding to the NCD1.2 monoclonal antibody.

• DOI: 10.1042/bcj20200674
• 发表时间: 2021-01-15
• 期刊: The Biochemical journal
• 作者: Uhrik L;Hernychova L;Muller P;Kalathiya U;Lisowska MM;Kocikowski M;Parys M;Faktor J;Nekulova M;Nortcliffe C;Zatloukalova P;Ruetgen B;Fahraeus R;Ball KL;Argyle DJ;Vojtesek B;Hupp TR
• 通讯作者: Hupp TR

The Development of a Recombinant scFv Monoclonal Antibody Targeting Canine CD20 for Use in Comparative Medicine.

• DOI: 10.1371/journal.pone.0148366
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Jain S;Aresu L;Comazzi S;Shi J;Worrall E;Clayton J;Humphries W;Hemmington S;Davis P;Murray E;Limeneh AA;Ball K;Ruckova E;Muller P;Vojtesek B;Fahraeus R;Argyle D;Hupp TR
• 通讯作者: Hupp TR

Quantitative proteomic profiling of pleomorphic human sarcoma identifies CLIC1 as a dominant pro-oncogenic receptor expressed in diverse sarcoma types.

• DOI: 10.1021/pr4010713
• 发表时间: 2014-04
• 期刊: Journal of proteome research
• 影响因子: 4.4
• 作者: E. Murray;L. Hernychova;M. Scigelová;J. Ho;M. Nekulová;J. R. O'Neill;R. Nenutil;K. Veselý;S. R. Dundas;C. Dhaliwal;H. Henderson;R. L. Hayward;D. Salter;B. Vojtěšek;T. Hupp