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（由申请人提供）：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分特性、肿瘤内高静水压和转移性肿瘤接受的低血流量。这些因素的结合限制了将大量治疗剂输送到肿瘤中。巨噬细胞的肿瘤归巢已被充分记录，并且最近的进展使得能够在体外产生可自我更新的、同基因的和完全分化的正常巨噬细胞（SSM）。我们建议通过基因插入或载药纳米颗粒（脂质体）从 BALB/c 小鼠中生成并修改 SSM，然后将“武装”SSM 重新注射回患有原位 4T1 乳腺肿瘤的 BALB/c 小鼠体内。我们将研究影响迁移到 4T1-luc 原发性肿瘤以及乳腺脂肪垫远端转移部位（如脑或肺）的 SSM 剂量百分比的因素，这些因素是使用荧光素酶报告基因在4T1线。我们将在 SSM 中装载维持巨噬细胞 M1 表型的药物，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，并确定肿瘤驻留巨噬细胞是否可以通过 SSM 分泌的 IL12 和干扰素 γ 恢复到 M1 表型。最后，我们将确定“武装”SSM 对 BALB/c 小鼠中 4T1 肿瘤的抗肿瘤活性。将积极追求三个具体目标。目标 1. 设计脂质体配方和培养条件，以便在体外高效地将纳米颗粒装载到 RAW309 Cr1 细胞中作为巨噬细胞原型。评估 4T1 小鼠乳腺肿瘤模型中脂质体负载巨噬细胞的比率、范围和肿瘤分布，作为注射巨噬细胞数量的函数。该目标的完成将指导目标 3 中的负荷、剂量和剂量频率。目标 2. 通过基因去除 c-Maf 和 MafB 功能，从 BALB/c 衍生的正常骨髓细胞中生成并表征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 修改 SSM。目标 3. 评估目标 2 中创建并负载药物纳米载体或诱导蛋白（如酵母胞嘧啶脱氨酶或干扰素 γ/IL12）的 SSM 在 BALB/c 4T1 乳腺癌模型中的肿瘤分布和治疗活性。我们的研究将对控制巨噬细胞运输到肿瘤的因素以及这种方式控制肿瘤生长和转移的潜力产生定量的了解。这项研究的成功可以促进基于自体巨噬细胞的新疗法的开发，从而大大增加治疗剂进入和遍及肿瘤的递送。它还可以提供一种有效的方法来重新教育肿瘤相关的 M2 型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，以根除目前可用方法无法实现的转移性肿瘤。

项目成果

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

氯膦酸盐可提高免疫功能小鼠中具有组成型活性 GMCSFR 的移植 Hoxb8 骨髓祖细胞的存活率。

• 发表时间: 2017-12-15
• 作者: Lee, Simon;Kivimäe, Saul;Szoka, Francis C
• 通讯作者: Szoka, Francis C

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

基于巨噬细胞的细胞疗法：漫长而曲折的道路。

• 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