Syngeneic Macrophages for Personalized Cancer Therapy

用于个性化癌症治疗的同基因巨噬细胞

• 批准号: 8787458
• 负责人: FRANCIS C. SZOKA
• 金额: $ 17.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787458
• 关键词: 4T1; Affect; Animals; Antibodies; Autologous; Back; Blood flow; Bone Marrow; Bone Marrow Cells; Brain; Breast Cancer Model; Cell Therapy; Cells; Cellular biology; Characteristics; Cytosine deaminase; Development; Distal; Dose; Drug Delivery Systems; Drug Formulations; Engineering; Environment; Excision; Fatty acid glycerol esters; Frequencies; Genes; Genetic; Goals; Health; Homing; Hydrostatic Pressure; In Vitro; Inbred BALB C Mice; Interferon Type II; Interleukin-12; Liposomes; Luciferases; Lung; Mammary Neoplasms; Mammary gland; Methods; Modality; Mus; Neoplasm Metastasis; Patients; Penetration; Pharmaceutical Preparations; Phenotype; Primary Neoplasm; Procedures; Production; Property; Proteins; Reporter; Research; Site; Solid Neoplasm; Testing; Therapeutic; Therapeutic Agents; Time; Tumor Tissue; Yeasts; arm; cancer therapy; chemotherapy; cytotoxic; extracellular; functional loss; improved; macrophage; nanocarrier; nanoparticle; personalized cancer therapy; protein expression; prototype; self-renewal; success; therapeutic protein; trafficking; transcription factor; tumor; tumor eradication; tumor growth; yeast protein

DESCRIPTION (provided by applicant): A number of barriers impede anti-tumor activity of drugs, antibodies, and nanoparticles including: sieving properties of the extracellular tumor matrix, high intra-tumoral hydrostatic pressures and the low volume of blood flow that metastatic tumors receive. The combination of these factors restrict the delivery of high quantities of therapeutic agents into tumors. The tumor homing of macrophages is well documented and recent advances enable the ex-vivo production of self-renewable, syngeneic and fully differentiated normal macrophages (SSM). We propose to generate and then modify SSM from BALB/c mice via gene insertions or drug-loaded nanoparticles (liposomes) and then re-inject the "armed" SSM back into BALB/c mice with orthotopic 4T1 breast tumors. We will investigate factors that affect the time course and percent of the SSM dose that migrates into the 4T1-luc primary tumor as well as into metastatic sites distal to the mammary fat pad such as the brain or lung, identified using the luciferase reporter in the 4T1 line. We will load agents into the SSM that maintain macrophages in the M1 phenotype to enhance their anti-tumor activity. We will investigate the effect of the SSM and the agents they deliver on the phenotype of the tumor resident macrophages and determine if tumor resident macrophages can be returned to a M1 phenotype by IL12 and Interferon gamma secreted by the SSM. Finally, we will determine the anti-tumor activity of "armed" SSM against the 4T1 tumor in BALB/c mice. Three specific aims will be aggressively pursued. Aim 1. Devise liposome formulations and culture conditions for highly efficient in vitro loading of nanoparticles into RAW309 Cr1 cells as a macrophage prototype. Evaluate the rate, extent and tumor distribution of liposome-loaded macrophages in the 4T1 murine breast tumor model as a function of the number of macrophages injected. Completion of this aim will guide the loading, dosing and frequency of doses in aim 3. Aim 2. Generate and characterize self-renewing syngeneic macrophages from BALB/c derived normal bone marrow cells by genetic removal of c-Maf and MafB function. We will also modify the SSM with a red reporter protein and yeast cytosine deaminase or interferon gamma/IL12. Aim 3. Evaluate the tumor distribution and therapeutic activity of SSM created in aim 2 and loaded with drug nanocarriers or inducible proteins such as yeast cytosine deaminase, or Interferon gamma/IL12 in BALB/c 4T1 breast cancer model. Our studies will generate a quantitative understanding of the factors that control the trafficking of macrophages into tumors as well as the potential for this modality to control tumor growth and metastases. Success in this research could enable the development of new autologous macrophage cell-based therapies that greatly increase therapeutic agent delivery into and throughout tumors. It could also provide an effective means to re-educate tumor-associated type M2 macrophages to become tumor killers. If effective, "armed" SSM could also provide a personalized cancer treatment to eradicate metastatic tumors that cannot be achieved by currently available approaches.

描述(申请人提供)：许多障碍阻碍药物、抗体和纳米颗粒的抗肿瘤活性，包括：细胞外肿瘤基质的筛选特性、高肿瘤内静水压力和转移性肿瘤获得的低血流量。这些因素的结合限制了向肿瘤中输送大量治疗剂。巨噬细胞的肿瘤归巢是有文献记载的，最近的研究进展使体外产生自我更新、同基因和完全分化的正常巨噬细胞(SSM)成为可能。我们建议通过基因插入或载药纳米粒(脂质体)从BALB/c小鼠身上产生SSM，然后对其进行修饰，然后将“武装”SSM重新注射到BALB/c小鼠体内，这些小鼠患有原位4T1乳腺肿瘤。我们将调查影响SSM剂量迁移到4T1-Luc原发肿瘤以及乳房脂肪垫远端转移部位(如脑或肺)的时间进程和百分比的因素，这是使用4T1线中的荧光素酶报告确定的。我们将向SSM中加载药物，使巨噬细胞保持M1表型，以增强其抗肿瘤活性。我们将研究SSM及其递送的药物对肿瘤驻留巨噬细胞表型的影响，并确定SSM分泌的IL12和干扰素γ能否使肿瘤驻留巨噬细胞恢复到M1表型。最后，我们将测定“武装”SSM对BALB/c小鼠4T1肿瘤的抗肿瘤活性。将积极追求三个具体目标。目的1.设计脂质体处方和培养条件，将纳米颗粒高效体外负载到RAW309 CR1细胞中作为巨噬细胞原型。评价在4T1小鼠乳腺肿瘤模型中，脂质体负载的巨噬细胞的比率、范围和肿瘤分布与注射的巨噬细胞数量的函数关系。这一目标的完成将指导目标3中剂量的加载、剂量和频率。目的2.通过遗传去除c-Maf和MafB功能，从BALB/c来源的正常骨髓细胞中产生并鉴定自我更新的同基因巨噬细胞。我们还将用红色报告蛋白和酵母胞嘧啶脱氨酶或干扰素γ/IL12修饰SSM。目的3.在BALB/c-4T1乳腺癌模型中，评价AIM-2制备的SSM负载药物纳米载体或诱导性蛋白如酵母胞嘧啶脱氨酶或干扰素-γ/IL-12的肿瘤分布和治疗活性。我们的研究将使我们对控制巨噬细胞进入肿瘤的因素以及这种方式控制肿瘤生长和转移的潜力有一个定量的了解。这项研究的成功可能使新的基于巨噬细胞的自体疗法的开发成为可能，这种疗法可以极大地增加治疗药物进入肿瘤和贯穿肿瘤的能力。它还可以提供一种有效的手段，重新教育肿瘤相关类型的M2巨噬细胞成为肿瘤杀手。如果有效，“武装”SSM还可以提供个性化的癌症治疗，以根除目前可用的方法无法实现的转移性肿瘤。

项目成果

Macrophage-based cell therapies: The long and winding road.

• DOI: 10.1016/j.jconrel.2016.07.018
• 发表时间: 2016-10-28
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: Lee S;Kivimäe S;Dolor A;Szoka FC
• 通讯作者: Szoka FC

Clodronate Improves Survival of Transplanted Hoxb8 Myeloid Progenitors with Constitutively Active GMCSFR in Immunocompetent Mice.

• DOI: 10.1016/j.omtm.2017.08.007
• 发表时间: 2017-12-15
• 期刊: Molecular therapy. Methods & clinical development
• 作者: Lee S;Kivimäe S;Szoka FC
• 通讯作者: Szoka FC