Active Immunity Targeted at PCSK9 for the Treatment of Hypercholesterolemia

描述（由申请人提供）：本提案的长期目标是设计和开发抑制/减少PCSK9作用的新策略。这是一个潜在的强大的新的治疗靶点来调节LDL胆固醇和减轻心血管疾病的负担。PCSK9 （Proprotein convertase subtilisin/ keexin type 9）是一个新发现的基因，它通过与LDL受体结合对LDL胆固醇水平产生深远的影响，并作为伴侣子介导LDL受体到溶酶体降解。人类研究表明，总PCSK9缺乏症的LDL胆固醇非常低（14-16 mg/dl），没有不良的临床后果。因此，PCSK9将是治疗高胆固醇血症的一个极好的靶点。此外，有研究表明他汀类药物增加PCSK9转录，导致血浆PCSK9水平升高。然后，他汀类药物和PCSK9抑制剂的联合治疗将提供更好的治疗来降低LDL胆固醇。在这个应用中，我们建议使用一种主动免疫治疗策略，通过产生一种疫苗来刺激免疫系统对抗PCSK9。蛋白质是独特的生物分子，它们展开和折叠，产生不同构象异构体的混合物。我们开发了一种技术的二硫化置乱允许可逆转换之间的原生和各种变性异构体。我们的初步结果表明，这些异构体诱导免疫原性对天然蛋白和降低血浆胆固醇水平。这是一种新颖的方法，利用二硫化物置乱产生Pcsk9的构象异构体（X- Pcsk9）来诱导对天然Pcsk9的免疫原性。这一过程将阻断PCSK9与LDL受体的相互作用，从而降低LDL胆固醇，从而治疗高胆固醇血症。此外，针对氧化LDL或载脂蛋白B肽的动脉粥样硬化疫苗也取得了一定的成功。因此，开发针对PCSK9的疫苗来调节LDL胆固醇的建议是一种令人兴奋和可行的方法。在本应用中，我们将生成和生产一组X-Pcsk9异构体疫苗，它们将与天然PCSK9交叉反应。我们建议评估这些疫苗在体内降低胆固醇水平和调节动脉粥样硬化发展方面的效力。总之，本研究将为降低LDL水平和减轻心血管疾病负担提供新的治疗方法。

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to design and develop novel strategies to inhibit/reduce PCSK9 action. This is a potential powerful new therapeutic target to regulate LDL cholesterol and reduce the burden of cardiovascular disease. PCSK9 (Proprotein convertase subtilisin/kexin type 9) is a newly discovered gene that has a profound effect on LDL cholesterol levels by binding to LDL receptor and acts as a chaperon, mediates LDL receptor to lysosome for degradation. Studies in Humans demonstrated that total PCSK9 deficiency had very low LDL cholesterol (14-16 mg/dl) with no adverse clinical consequences. Thus, PCSK9 would be an excellent target for the treatment of hypercholesterolemia. Moreover, it has been shown that statins increase PCSK9 transcription, resulting in higher plasma PCSK9 levels. Then, a combination treatment of statins and PCSK9 inhibitor(s) would provide a better therapy to reduce LDL cholesterol. In this application, we propose to use an active immunotherapy strategy by generating a vaccine that stimulates the immune system against PCSK9. Proteins are unique bio-molecules, they unfold and refold, generate a mixture of diverse conformational isomers. We developed a technique of disulfide scrambling permits reversible conversion between the native and diverse denatured isomers. Our preliminary results have shown that these isomers induced immunogenicity towards native protein and decreased plasma cholesterol levels. This is a novel approach to use disulfide scrambling to generate conformational isomers of Pcsk9 (X- Pcsk9) to induce immunogenicity towards native Pcsk9. This process will block the interaction of PCSK9 and LDL receptor to decrease LDL cholesterol for the treatment of hypercholesterolemia. Moreover, Vaccine for atherosclerosis towards oxidized LDL or apolipoprotein B peptide has shown some success. Thus, the proposal of developing vaccine against PCSK9 to regulate LDL cholesterol is an exciting and feasible approach In this application, we will generate and produce a panel of X-Pcsk9 isomers vaccines, which will cross- react with native Pcsk9. We propose to evaluate the potency of these vaccines on decreasing cholesterol levels as well as modulating atherosclerosis development in vivo. In summary, this study will provide novel therapeutic treatment for decreasing LDL levels and reducing the burden of cardiovascular disease.