Semi-targeted Immuno-Therapeutic Approach for Treatment of Obesity

描述(申请人提供)：在美国和欧洲，肥胖被认为是头号健康问题，影响10%到30%的成年人口。肥胖的人更有可能患上各种继发性疾病，包括糖尿病、心脏问题、免疫缺陷、某些类型的癌症和骨关节炎，而且他们比非肥胖的人死得更早。肥胖是由于过多的脂肪细胞(脂肪细胞)的发育和全身现有脂肪细胞的增大造成的。减少卡路里摄入量和增加体力活动往往是不可持续的，而且几乎没有安全的替代疗法。没有一种治疗方法可以利用现代分子医学在免疫治疗方面的进步。单抗的治疗应用提供了一种针对特定组织的靶向治疗方法。然而，尽管大量研究表明，针对脂肪细胞质膜抗原的单抗可用于抑制动物模型中的脂肪发育，但目前还没有用于治疗或诊断人类肥胖症的单抗。因此，这个第一阶段项目的直接目标是使用Abeome的快速高效的DISH(直接选择杂交瘤)技术来分离人内脏前脂肪细胞和脂肪细胞表面的一组针对PMAS的单抗。我们的长期目标是开发免疫疗法，提供各种针对脂肪细胞的结果，例如刺激白色脂肪细胞向棕色脂肪细胞的转分化，抑制脂肪生成，或诱导细胞凋亡。我们第一阶段的具体目标如下。1.用人内脏前脂肪细胞免疫20只小鼠，筛选出10只抗人内脏前脂肪细胞PMAs抗体效价最高的小鼠。#2.使用Abeome的DISH-PMA方案分离至少100株产生人类内脏前脂肪细胞和脂肪细胞抗体的杂交瘤细胞。#3.使用全细胞ELISA和免疫荧光显微镜(IFM)鉴定20种单抗试剂，它们与人内脏前脂肪细胞和脂肪细胞表面的PMAs反应，但不与人脂肪细胞来源的干细胞(ADSC)、人间充质干细胞(HMSC)、人皮下前脂肪细胞或脂肪细胞或来自其他人体组织的对照细胞反应。Abeome及其学术合作者计划提交一份第二阶段提案，进一步开发脂肪细胞特异性单抗作为治疗、诊断和研究试剂。我们将确定特定的人类单抗，这些单抗可以稳定地偶联到药物递送剂上，并进行体外测试。最合适的给药方法将在体内进行测试，以提供一种刺激白色脂肪细胞向棕色脂肪细胞转分化的化合物，或者一种专门抑制脂肪生成或诱导细胞凋亡的化合物。然后将在适当的动物模型中测试抗体靶向药物传递。这些研究中产生的数据将作为这种免疫疗法治疗肥胖症的临床评估的基础。

DESCRIPTION (provided by applicant): Obesity is considered the number one health problem in the US and Europe, affecting 10% to 30% of adult populations. Obese individuals are more likely to acquire a variety of secondary diseases including diabetes, heart problems, immune deficiencies, some types of cancer, and osteoarthritis and they die younger than non-obese individuals. Obesity results from the development of too many fat cells (adipocytes) and the enlargement of existing fat cells throughout the body. Reducing caloric intake and increasing physical activity are often not sustainable and there are few safe alternative treatments. There are no treatments that take advantage of modern molecular medicine's advances in immunotherapy. Therapeutic applications of monoclonal antibodies (mAbs) offer a way to target treatments to specific tissues. However, despite substantial work showing that mAbs to adipocyte plasma membrane antigens (PMAs) may be used to suppress fat development in animal models, there are currently no mAbs for either therapeutic or diagnostic applications in human obesity. Thus, the immediate goal for this Phase I project is to use Abeome's rapid and efficient DiSH" (Direct Selection of Hybridomas) technology to isolate a battery of mAbs specific for PMAs on the surface of human visceral preadipocytes and adipocytes. Our long-term goal is to develop immuno-therapeutics that offer a variety of adipocyte- targeted outcomes, such as stimulating transdifferentiation of white adipocytes to brown adipocytes, inhibiting adipogenesis, or inducing apoptosis. Our Phase I Specific Aims are as follows. #1. Immunize 20 mice with human visceral preadipocytes and identify 10 mice with the best antibody titers to PMAs of human visceral preadipocytes and adipocytes. #2. Isolate at least one hundred hybridomas making antibodies to human visceral preadipocytes and adipocytes using Abeome's DiSH-PMA protocol. #3. Using whole cell ELISAs and immunofluorescence microscopy (IFM) identify 20 mAb reagents that react with surface PMAs on human visceral preadipocytes and adipocytes but not with human adipocyte derived stem cells (ADSC), human mesenchymal stem cells (hMSC), human subcutaneous preadipocytes or adipocytes, or control cells from other human tissues. Abeome and its academic collaborators plan to submit a Phase II proposal to further develop adipocyte- specific mAbs as therapeutic, diagnostic and research agents. We will identify specific human mAbs that can be stably coupled to drug delivery agents and tested in vitro. The most appropriate drug delivery method will be tested in vivo to deliver a compound that stimulates transdifferentiation of white adipocytes to brown-like adipocytes or a compound that specifically inhibits adipogenesis or that induces apoptosis. Antibody targeted drug delivery will then be tested in appropriate animal models. The data generated in these studies will serve as the basis for clinical evaluation of this immuno-therapeutic approach to treating obesity.