Tumor Microenvironment in Cancer Progression

癌症进展中的肿瘤微环境

• 批准号: 10926185
• 负责人: Rosandra Kaplan
• 金额: $ 119.07万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10926185
• 关键词: Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Adhesions; Adjuvant; Adrenocortical carcinoma; Adult; Area; Blood Vessels; Bone Marrow; Breast; Breast Carcinoma; CSF1R gene; Cell Communication; Cell Compartmentation; Cell Line; Cell Survival; Cell Therapy; Cells; Cellular Assay; Cessation of life; Characteristics; Childhood; Childhood Lymphoma; Childhood Rhabdomyosarcoma; Circulation; Clinic; Clinical; Colon; Cyclophosphamide; Data; Development; Disease; Distant; Dose; Dysmyelopoietic Syndromes; Endothelial Cells; Environment; Equilibrium; Event; Ewings sarcoma; Extracellular Matrix; FLT3 gene; Fibroblasts; Genes; Genetic Engineering; Glioma; Goals; Growth; Hematopoietic; Hematopoietic Stem Cell Mobilization; Hematopoietic stem cells; High-Risk Cancer; Human; ITGAM gene; Immune; Immune Evasion; Immunosuppression; Immunotherapy; In Vitro; Individual; Inflammation; Interleukin-12; Investigation; Lead; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of adrenal cortex; Malignant neoplasm of gastrointestinal tract; Malignant neoplasm of lung; Mediating; Mesenchymal; Metalloproteases; Metastatic Neoplasm to the Bone; Modeling; Molecular; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; Natural Killer Cells; Neoplasm Metastasis; Nerve; Neuroblastoma; Organ; Pathway interactions; Patients; Pediatric Neoplasm; Pericytes; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Play; Population; Pre-Clinical Model; Primary Neoplasm; Proliferating; Proteins; Recurrence; Refractory; Reproducibility; Rhabdomyosarcoma; Role; Sampling; Signal Transduction; Site; Soil; Solid Neoplasm; Stromal Cells; Supporting Cell; T cell response; T-Lymphocyte; Tissues; Translating; Tumor Immunity; Tumor Promotion; Tumor-Derived; Tumor-infiltrating immune cells; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors; Work; adolescent patient; angiogenesis; anticancer research; arginase; bone; bone scaffold; cancer therapy; cancer type; cellular engineering; combinatorial; design; exosome; fludarabine; genetic manipulation; hematopoietic stem cell niche; human tissue; in vivo; insight; lung Carcinoma; melanoma; metastatic process; mouse model; neoplastic cell; neurofibroma; novel; novel strategies; novel therapeutic intervention; osteosarcoma; pediatric patients; pre-clinical; prevent; programs; recruit; response; response to injury; sarcoma; small molecular inhibitor; stem cell biology; stem cell niche; stem cells; therapy design; therapy development; tissue injury; tissue repair; transcription factor; translational study; tumor; tumor growth; tumor microenvironment; tumor progression; tumor-immune system interactions

We have established that during primary tumor growth there is a formation of a niche environment in distant tissue sites during metastatic initiation. This pre-metastatic tissue has an influx of bone marrow-derived cells including populations of myeloid cells and overall expansion in myeloid cells that induce immunosuppressive of T and NK cell mediated anti-tumor immunity. These myeloid cells also provide factors such as matrix metalloproteases to remodel extracellular matrix and pro-growth and survival signals such as VEGF and arginase to support the colonizing disseminated tumor cells and allow for immune evasion. These sites are created as a systemic response to tumor progression and may be organ specific and activate tissue injury response programs in specific microenvironments. Using syngeneic cells lines that have a high spontaneous metastatic rate, we have identified unique changes within the bone marrow microenvironment that lead to mobilization of bone marrow-derived cells that are recruited to the pre-metastatic niche in multiple tumor models including E0771 breast carcinoma, F4 osteosarcoma, 76-9 and M3-9M pediatric rhabdomyosarcomas and B16 melanoma. Previously we have shown that CD11b myeloid cells expressed VEGFR1 in the pre-metastatic tissue. We have now discovered these cells are hematopoietic progenitor cells that become an immune suppressive myeloid population that alter the local immune environment favoring immune evasion similar to sanctuary sites in stem cell niches (Giles et al Cancer Research 2016). We have also identified the central role of immune suppression in creation of pre-metastatic and early metastatic niche and the means of reprogramming this immune suppression to limit metastasis (Kaczanowska et al Cell 2021). These cells are immune suppressive myeloid cells derived from mobilized bone marrow-derived hematopoietic stem and progenitor cells which play an integral role in regulating T cell responses (Giles et al Cancer Research 2016; Kaczanowska et al Cell 2021). We are able to manipulate metastatic progression by altering these unique bone marrow-derived cell enriched areas. We have new data demonstrating that the pre-metastatic niche has similar features to physiological stem cell niches in order to promote distant tumor cell survival. We have found that the localized tumor prior to established metastasis is activating the hematopoietic stem cell niche within the bone marrow and inducing proliferation of hematopoietic stem cells and mobilization of these cells into the circulation. We have found that there are changes that occur in the bone marrow microenvironment in response to tumor secreted factors that induce the myeloid skewing and expansion of hematopoietic progenitor cells that we have seen during tumor progression (Giles et al Cancer Research 2016). Targeting the skewing to prevent the expansion in hematopoietic progenitor cells and myeloid cells may be a way to reset this maladaptive response to a growing tumor and prevent metastatic progression. We have on-going investigations examining the small molecular inhibitor PLX3397 that targets CSF1R found on myeloid cells, cKit and FLT3-ITD which we have determined that when the drug is given in the adjuvant setting can limit metastatic progression in tumor bearing hosts. We have initiated and completed the Phase I dose escalation of PLX3397 in pediatric and adolescent patients with recurrent or refractory tumors (Boal et al Clinical Cancer Research). We have also developed a new cell therapy approach platform to reprogram these recruited immune suppressive myeloid cells by introducing genetically engineered myeloid cells (GEMys) that express IL12. These IL12 GEMys can reverse the immune suppression program in the pre-metastatic niche and lead to inhibition of metastatic progression and significantly prolong overall survival in highly metastatic murine models (Kaczanowska et al Cell 2021). IL12 GEMys when given following fludarabine and cyclophosphamide tumor bearing hosts lead to long term cures in these metastatic pre-clinical models (Kaczanowska et al Cell 2021). In addition to investigations into the recruited hematopoietic progenitor bone marrow derived cell populations that become immune suppressive cells in pre-metastatic sites, we continue to investigate the essential changes in stromal cells including pericytes, vascular cells and fibroblasts as well as the extracellular matrix in the pre-metastatic and metastatic niche. We have established several lineage tracing models to better track and characterize these stromal cell populations as well as genetically manipulate key genes within specific cell populations. Using these models, we can interrogate the function of specific proteins to these cells and their role in the metastatic process. A specific transcription factor KLF4 we have discovered is critical to mediating this stromal cell plasticity. These stromal cells that become activated create a distinct extracellular matrix that support disseminated tumor cell survival. We are currently investigating the role of tumor conditioned media and tumor derived exosomes in making local changes in the stromal cell compartment and matrix that provides the scaffolding for bone marrow-derived cells and are essential component of the pre-metastatic niche (Murgai et al Nat Med 2017). We have identified two critical cellular pathways in pre-metastatic niche formation related to inflammation and stem cell biology involving myeloid and stromal cells. Understanding the activation of these tissue repair pathways and inflammation- related pathways in the metastatic process are an active area of investigation. In addition to our existing stromal models, we have developed a new lineage tracing model to track mesenchymal cells that support nerves. These models are robust and show a new phenotype of mesenchymal nerve support cells that are found in bone and other sites and we have preliminary data to suggest a function in the pre-metastatic niche in bone during bone metastasis. Our current investigations reveal important components of the dysregulated microenvironments that occur during metastatic progression. Our novel cell therapy approach can provide a platform to locally rebalance these altered microenvironments and a potential new therapeutic approach to limit metastatic progression in pediatric and adult patients at high risk for cancer progression. We are developing multiple approaches to translate this new cell therapy platform into the clinic setting. We are also actively exploring which cargo are most effective to rebalance dysregulated microenvironments that occur during metastasis. As many cancers have a dense extracellular matrix and we determined that extracellular matrix remodeling is occurring in an enhanced fashion from stromal cell expansion. We are investigating modulation of extracellular matrix through cell therapy delivery of extracellular matrix remodeling proteins. More dense matrix is associated with malignant tissue. We aim to explore the impact of matrix remodeling on metastatic progression. We have developed robust cell engineering strategies, culture conditions and reproducible assays for cell therapy development. We continue to develop functional studies that help demonstrate human genetically engineered myeloid cell therapy (GEMys) can effectively deliver the cargo and that the cargo has the anticipated response in vitro and in preclinical in vivo studies. We are preforming extensive IND enabling studies. We are also developing correlative markers to explore response and microenvironmental changes in immunotherapy in solid tumors. These studies can not only provide insights into response but also development of potential combinatorial approaches to target diverse aspects of the tumor microenvironment.

我们已经确定，在原发性肿瘤生长过程中，在转移起始期间，远处组织部位会形成利基环境。这种转移前组织大量涌入骨髓源性细胞，包括骨髓细胞群，并且骨髓细胞整体扩张，诱导 T 细胞和 NK 细胞介导的抗肿瘤免疫的免疫抑制。这些骨髓细胞还提供基质金属蛋白酶等因子来重塑细胞外基质，并提供促生长和存活信号（例如 VEGF 和精氨酸酶）以支持定植播散性肿瘤细胞并允许免疫逃避。这些位点是作为对肿瘤进展的系统反应而产生的，并且可能是器官特异性的，并在特定的微环境中激活组织损伤反应程序。使用具有高自发转移率的同基因细胞系，我们发现了骨髓微环境内的独特变化，这些变化导致骨髓衍生细胞被动员到多种肿瘤模型中的转移前生态位，包括E0771乳腺癌、 F4 骨肉瘤、76-9 和 M3-9M 儿童横纹肌肉瘤和 B16 黑色素瘤。之前我们已经证明 CD11b 骨髓细胞在转移前组织中表达 VEGFR1。我们现在发现这些细胞是造血祖细胞，它们成为免疫抑制性骨髓细胞群，改变局部免疫环境，有利于免疫逃避，类似于干细胞生态位中的避难所（Giles et al Cancer Research 2016）。我们还确定了免疫抑制在转移前和早期转移生态位创建中的核心作用，以及重新编程这种免疫抑制以限制转移的方法（Kaczanowska et al Cell 2021）。这些细胞是源自动员的骨髓源性造血干细胞和祖细胞的免疫抑制性骨髓细胞，在调节 T 细胞反应中发挥着不可或缺的作用（Giles 等人 Cancer Research 2016；Kaczanowska 等人 Cell 2021）。我们能够通过改变这些独特的骨髓来源细胞富集区域来控制转移进展。我们有新的数据表明，转移前的微环境与生理干细胞微环境具有相似的特征，以促进远处肿瘤细胞的存活。我们发现，在发生转移之前的局部肿瘤正在激活骨髓内的造血干细胞生态位，并诱导造血干细胞增殖并将这些细胞动员到循环中。我们发现，在肿瘤进展过程中，我们已经看到，肿瘤分泌因子会导致骨髓微环境发生变化，这些变化会诱导造血祖细胞的骨髓偏斜和扩张（Giles 等癌症研究 2016）。针对这种倾斜以防止造血祖细胞和骨髓细胞的扩张可能是重置这种对生长肿瘤的适应不良反应并防止转移进展的一种方法。我们正在进行研究，检查针对骨髓细胞、cKit 和 FLT3-ITD 上发现的 CSF1R 的小分子抑制剂 PLX3397，我们已经确定，当在辅助环境中给予该药物时，可以限制荷瘤宿主的转移进展。我们已经启动并完成了 PLX3397 在患有复发性或难治性肿瘤的儿童和青少年患者中的 I 期剂量递增（Boal 等人临床癌症研究）。我们还开发了一种新的细胞治疗方法平台，通过引入表达 IL12 的基因工程骨髓细胞 (GEMys) 来重新编程这些招募的免疫抑制性骨髓细胞。这些 IL12 GEMys 可以逆转转移前生态位中的免疫抑制程序，抑制转移进展，并显着延长高度转移小鼠模型的总体生存期 (Kaczanowska et al Cell 2021)。当氟达拉滨和环磷酰胺肿瘤宿主给予 IL12 GEMys 时，可在这些转移性临床前模型中实现长期治愈 (Kaczanowska et al Cell 2021)。除了研究招募的造血祖细胞骨髓来源的细胞群在转移前成为免疫抑制细胞外，我们还继续研究基质细胞（包括周细胞、血管细胞和成纤维细胞）以及细胞外基质的基本变化。转移前和转移生态位。我们建立了几种谱系追踪模型，以更好地追踪和表征这些基质细胞群，并对特定细胞群内的关键基因进行遗传操作。使用这些模型，我们可以探究特定蛋白质对这些细胞的功能及其在转移过程中的作用。我们发现的一种特定转录因子 KLF4 对于介导这种基质细胞可塑性至关重要。这些被激活的基质细胞产生独特的细胞外基质，支持播散性肿瘤细胞的存活。我们目前正在研究肿瘤条件培养基和肿瘤来源的外泌体在基质细胞区室和基质发生局部变化中的作用，基质细胞区室和基质为骨髓来源的细胞提供支架，并且是转移前生态位的重要组成部分（Murgai et al Nat医学2017）。我们已经确定了转移前生态位形成中与炎症和涉及骨髓和基质细胞的干细胞生物学相关的两条关键细胞途径。了解转移过程中这些组织修复途径和炎症相关途径的激活是一个活跃的研究领域。除了我们现有的基质模型之外，我们还开发了一种新的谱系追踪模型来追踪支持神经的间充质细胞。这些模型是稳健的，并显示了在骨和其他部位发现的间充质神经支持细胞的新表型，我们有初步数据表明骨转移期间骨中转移前生态位的功能。我们目前的研究揭示了转移进展过程中发生的失调微环境的重要组成部分。我们的新型细胞治疗方法可以提供一个平台来局部重新平衡这些改变的微环境，并提供一种潜在的新治疗方法来限制癌症进展高风险的儿科和成人患者的转移进展。我们正在开发多种方法将这种新的细胞治疗平台转化为临床环境。我们还在积极探索哪些货物最有效地重新平衡转移过程中出现的失调微环境。由于许多癌症具有致密的细胞外基质，我们确定细胞外基质重塑因基质细胞扩张而以增强的方式发生。我们正在研究通过细胞疗法递送细胞外基质重塑蛋白来调节细胞外基质。更致密的基质与恶性组织相关。我们的目的是探讨基质重塑对转移进展的影响。我们为细胞疗法的开发开发了强大的细胞工程策略、培养条件和可重复的检测方法。我们继续开展功能研究，帮助证明人类基因工程骨髓细胞疗法 (GEMys) 可以有效地递送货物，并且货物在体外和临床前体内研究中具有预期的反应。我们正在开展广泛的 IND 授权研究。我们还在开发相关标记物来探索实体瘤免疫治疗的反应和微环境变化。这些研究不仅可以提供对反应的见解，还可以开发针对肿瘤微环境不同方面的潜在组合方法。

项目成果

Infectious complications of CAR T-cell therapy across novel antigen targets in the first 30 days.

CAR T 细胞疗法在前 30 天内跨新抗原靶标的感染并发症。

• 发表时间: 2021
• 影响因子: 7.5
• 作者: Mikkilineni, Lekha;Yates, Bonnie;Steinberg, Seth M;Shahani, Shilpa A;Molina, John C;Palmore, Tara;Lee, Daniel W;Kaplan, Rosandra N;Mackall, Crystal L;Fry, Terry J;Gea;Jerussi, Theresa;Nussenblatt, Veronique;Kochenderfer, Jame
• 通讯作者: Kochenderfer, Jame

Corrigendum: Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET.

勘误表：黑色素瘤外泌体通过 MET 将骨髓祖细胞培养为促转移表型。

• 发表时间: 2016-12-06
• 影响因子: 82.9
• 作者: Peinado, Héctor;Alecˇković, Maša;Lavotshkin, Simon;Matei, Irina;Costa;Moreno;Hergueta;Williams, Caitlin;García;Ghajar, Cyrus M;Nitadori;Hoffman, Caitlin;Badal, Karen;Gar
• 通讯作者: Gar

Tumor Mutation Burden, Expressed Neoantigens and the Immune Microenvironment in Diffuse Gliomas.

弥漫性胶质瘤中的肿瘤突变负担、表达的新抗原和免疫微环境。

• 发表时间: 2021-12-03
• 影响因子: 5.2
• 作者: Yu, Guangyang;Pang, Ying;Merchant, Mythili;Kesserwan, Chimene;Gangalapudi, Vineela;Abdelmaksoud, Abdalla;Ranjan, Alice;Kim, Olga;Wei, Jun S;Chou, Hsien;Wen, Xinyu;Sindiri, Sivasish;Song, Young K;Xi, Liqiang;Kaplan, Rosandra N;Armstrong
• 通讯作者: Armstrong

4-1BB costimulation ameliorates T cell exhaustion induced by tonic signaling of chimeric antigen receptors.

4-1BB 共刺激可改善嵌合抗原受体的强直信号传导诱导的 T 细胞耗竭。

• 发表时间: 2015-06
• 影响因子: 82.9
• 作者: Long, Adrienne H;Haso, Waleed M;Shern, Jack F;Wanhainen, Kelsey M;Murgai, Meera;Ingaramo, Maria;Smith, Jillian P;Walker, Alec J;Kohler, M Eric;Venkateshwara, Vikas R;Kaplan, Rosandra N;Patterson, George H;Fry, Terry J;Orentas, Rimas J;Mackal
• 通讯作者: Mackal

The functional interplay between systemic cancer and the hematopoietic stem cell niche.

系统性癌症与造血干细胞生态位之间的功能相互作用。

• DOI: 10.1016/j.pharmthera.2016.09.006
• 发表时间: 2016-12
• 期刊: Pharmacology & therapeutics
• 影响因子: 13.5
• 作者: Giles, Amber J.;Chien, Christopher D.;Reid, Caitlin M.;Fry, Terry J.;Park, Deric M.;Kaplan, Rosandra N.;Gilbert, Mark R.
• 通讯作者: Gilbert, Mark R.