Antibody-interferon fusion proteins for treatment of B-cell malignancies

用于治疗 B 细胞恶性肿瘤的抗体-干扰素融合蛋白

基本信息

批准号：
8657920
负责人：
Sherie L Morrison
金额：
$ 31万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8657920
关键词：
1-Phosphatidylinositol 3-Kinase Amino Acid Sequence Antibodies Apoptosis Autophagocytosis B lymphoid malignancy B-Cell Lymphomas B-Cell NonHodgkins Lymphoma Bioinformatics CD20 Antigens Caspase Cell Line Cell Proliferation Cell Survival Cell-Mediated Cytolysis Cells Characteristics Chimeric Proteins Chinese Hamster Ovary Cell Cleaved cell Clinical Complement Data Databases Disease Engineering Exhibits Goals Growth Health Histology Human Immune response Immune system Immunocompetent In Vitro Inbred BALB C Mice Inbred C3H Mice Individual Inhibition of Apoptosis Interferon Type I Interferon-alpha Interferon-beta Interferons Lymphoma MS4A1 gene Malignant Neoplasms Measures Mediating Modeling Monoclonal Antibodies Multiple Myeloma Mus Pathway interactions Peptide Sequence Determination Peptides Prostate Proteins Recruitment Activity Renal Cell Carcinoma Research Residual Cancers Resistance SCID Mice Sirolimus Site Specimen Surface Technology Tertiary Protein Structure Therapeutic Agents Toxic effect Transgenic Mice United States Xenograft procedure antibody-dependent cell cytotoxicity antitumor agent cancer cell cancer site cell growth clinically relevant in vitro activity in vivo inhibitor/antagonist mTOR Inhibitor melanoma neoplastic cell novel therapeutics programs therapeutic protein tumor

项目摘要

DESCRIPTION (provided by applicant): The type I interferons (IFNa and IFN¿) are potent regulators of cell growth with inhibitory effects against many human cancers. However, their clinical use has been limited by inability to achieve effective concentrations of IFN at sites of tumor without systemic toxicity. Our goal is to overcome this limitation using the tumor-targeting ability of monoclonal antibodies to carry IFNs directly to cancer sites. We hypothesize that antibody-IFN fusion proteins could be highly effective cancer therapeutic agents, selectively localizing IFN to sites of tumor. We have recently succeeded in producing fusion proteins targeting the CD20 antigen expressed on the surface of B cell non-Hodgkin lymphomas. Anti-CD20-IFNa fusion proteins containing murine or human IFNa exhibited potent anti-tumor effects against a human CD20+ murine lymphoma and human B cell lymphoma both in vitro and in vivo. We now propose to further characterize and optimize these IFN fusion proteins and to extend our studies to include fusion with IFN¿, which is up to 10-fold more potent than IFNa. To achieve this, we propose the following Specific Aims: Aim 1: Characterize the efficacy of anti-CD20 fused with murine IFNa (mIFNa) or murine IFN¿ (mIFN¿) against mouse lymphomas in vitro and in vivo and determine their mechanisms of action. A. Characterize the in vitro activity of fusion proteins against two murine B cell lymphoma cell lines engineered to express human CD20, including their ability to inhibit tumor cell proliferation, induce apoptosis, and mediate antibody-dependent cellular cytotoxicity (ADCC) and complement dependent cellular cytotoxicity (CDC). B. Characterize the in vivo efficacy of anti-CD20-mIFNa and anti-CD20-mIFN¿ against 38C13-huCD20 in C3H mice and A20-huCD20 in BALB/c huCD20-transgenic mice. Aim 2: Evaluate the efficacy of anti-CD20 fused with human IFNa (hIFNa) against human lymphomas in vitro and in vivo and determine the mechanism(s) of action. A. Using human B cell lymphoma cell lines representing various histologies, characterize the in vitro activity of anti-CD20-hIFNa as in Aim 1. B. Evaluate the in vivo efficacy of anti-CD20-hIFNa against human B cell lymphoma xenografts in SCID mice. C. Measure the activity of anti-CD20-hIFNa against primary human B cell lymphoma specimens in vitro. Aim 3: Construct anti-CD20 fusions with human IFN¿ in an attempt to increase anti-tumor potency. A. Explore the use of alternative peptide linkers to optimize the stability and IFN activity of the fusion protein. B. Characterize the most promising anti-CD20-hIFN¿ fusion proteins. Aim 4: Explore the possibility of further enhancing anti-CD20-hIFNa/¿ efficacy using clinically-relevant pharmacologic inhibitors of lymphoma cell survival pathways. A. Evaluate the in vitro efficacy of the fusion proteins against a panel of lymphoma cell lines and primary cells when used in combination with inhibitors of mTOR (rapamycin or temsirolimus) or PI3 kinase d (CAL-101). B. Evaluate the in vivo efficacy of the fusion proteins against human lymphoma xenografts in combination with inhibitors of mTOR (temsirolimus) or PI3 kinase d (CAL-120).

描述（由适用提供）：I型干扰素（IFNA和IFN¿）是细胞生长的潜在调节剂，对许多人类癌症具有抑制作用。但是，它们的临床用途受到无法在没有全身毒性的肿瘤部位实现有效浓度IFN的限制。我们的目标是利用单克隆抗体将IFN直接运送到癌症部位的肿瘤靶向能力来克服这一局限性。我们假设抗体-IFN融合蛋白可能是高效的癌症治疗剂，有选择地将IFN定位于肿瘤部位。最近，我们成功地产生了靶向在B细胞非霍奇金淋巴瘤表面表达的CD20抗原的融合蛋白。抗CD20-IFNA融合蛋白含有鼠或人IFNA暴露于人CD20+鼠淋巴瘤和人类B细胞淋巴瘤的有效抗肿瘤作用均在体外和体内。现在，我们建议进一步表征和优化这些IFN融合蛋白，并扩展我们的研究，以与IFN”进行融合，该蛋白比IFNA的潜力高10倍。为此，我们提出以下特定目的：目标1：表征与小鼠IFNA（mifna）或鼠ifn¿（mifn？）抗CD20的效率，以在体外和体内对小鼠淋巴瘤进行针对小鼠淋巴瘤的效率，并确定其作用机理。 A.表征融合蛋白对两种鼠类B细胞淋巴瘤细胞系的体外活性，这些细胞系设计为表达人CD20，包括它们抑制肿瘤细胞增殖的能力，诱导凋亡，并介导抗体依赖性细胞胞毒性（ADCC）和补体依赖性细胞胞毒性（CDC）。 B.表征抗CD20-MIFNA和抗CD20-MIFN的体内效率与C3H小鼠中38C13-HUCD20的体内效率和BALB/C HUCD20- TRANSGENIC小鼠中的38C13-HUCD20和A20-HUCD20。 AIM 2：评估与人IFNA（HIFNA）在体外和体内对人淋巴瘤融合的抗CD20的效率，并确定作用的机制。 A.使用代表各种组织学的人B细胞淋巴瘤细胞系，表征抗CD20-HIFNA的体外活性，如AIM1。B。评估抗CD20-HIFNA对SCID小鼠中人类B细胞淋巴瘤型的体内疗效。 C.在体外测量抗CD20-HIFNA对原代人B细胞淋巴瘤标本的活性。 AIM 3：与人IFN”构建抗CD20融合，以提高抗肿瘤效力。答：探索替代肽接头的使用以优化融合蛋白的稳定性和IFN活性。 B.表征最有希望的抗CD20-HIFN“融合蛋白。 AIM 4：探索使用淋巴瘤细胞存活途径的临床上相关的药物抑制剂进一步增强抗CD20-HIFNA/»有效性的可能性。答：当与MTOR（雷帕霉素或temsirolimus）或PI3激酶D（Cal-101）结合使用时，评估融合蛋白对淋巴瘤细胞系和原代细胞的体外效率。 B.评估融合蛋白针对人淋巴瘤异种移植物的体内效率与MTOR抑制剂（Temsirolimus）或PI3激酶D（Cal-120）结合使用。