Engineered CAR-Macrophages as Adoptive Immunotherapies for Solid Tumors

Cellular immunotherapies represent a promising approach for the treatment of cancer. Engineered adoptive cell therapies redirect and augment a leukocyte’s inherent ability to mount an immune response by introducing novel anti-tumor capabilities and targeting moieties. A prominent example of this approach is the use of T cells engineered to express chimeric antigen receptors (CARs), which have demonstrated significant efficacy against some hematologic malignancies. Despite increasingly sophisticated strategies to harness immune cell function, efficacy against solid tumors has remained elusive for adoptive cell therapies. Amongst cell types used in immunotherapies, however, macrophages have recently emerged as prominent candidates for the treatment of solid tumors. In this review, we discuss the use of monocytes and macrophages as adoptive cell therapies. Macrophages are innate immune cells that are intrinsically equipped with broad therapeutic effector functions, including active trafficking to tumor sites, direct tumor phagocytosis, activation of the tumor microenvironment and professional antigen presentation. We focus on engineering strategies for manipulating macrophages, with a specific focus on CAR macrophages (CAR-M). We highlight CAR design for macrophages, the production of CAR-M for adoptive cell transfer, and clinical considerations for their use in treating solid malignancies. We then outline recent progress and results in applying CAR-M as immunotherapies. The recent development of engineered macrophage-based therapies holds promise as a key weapon in the immune cell therapy armamentarium.

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Administration of cardiac mesenchymal cells modulates innate immunity in the acute phase of myocardial infarction in mice

Scientific Reports ◽

10.1038/s41598-020-71580-z ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Yi Kang ◽

Marjan Nasr ◽

Yiru Guo ◽

Shizuka Uchida ◽

Tyler Weirick ◽

...

Keyword(s):

Myocardial Infarction ◽

Innate Immunity ◽

Mechanism Of Action ◽

Cell Activation ◽

Cell Function ◽

Immune Cell ◽

Mesenchymal Cell ◽

Cell Types

Abstract Although cardiac mesenchymal cell (CMC) therapy mitigates post-infarct cardiac dysfunction, the underlying mechanisms remain unidentified. It is acknowledged that donor cells are neither appreciably retained nor meaningfully contribute to tissue regeneration—suggesting a paracrine-mediated mechanism of action. As the immune system is inextricably linked to wound healing/remodeling in the ischemically injured heart, the reparative actions of CMCs may be attributed to their immunoregulatory properties. The current study evaluated the consequences of CMC administration on post myocardial infarction (MI) immune responses in vivo and paracrine-mediated immune cell function in vitro. CMC administration preferentially elicited the recruitment of cell types associated with innate immunity (e.g., monocytes/macrophages and neutrophils). CMC paracrine signaling assays revealed enhancement in innate immune cell chemoattraction, survival, and phagocytosis, and diminished pro-inflammatory immune cell activation; data that identifies and catalogues fundamental immunomodulatory properties of CMCs, which have broad implications regarding the mechanism of action of CMCs in cardiac repair.

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Chimeric antigen receptors that trigger phagocytosis

eLife ◽

10.7554/elife.36688 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 43

Author(s):

Meghan A Morrissey ◽

Adam P Williamson ◽

Adriana M Steinbach ◽

Edward W Roberts ◽

Nadja Kern ◽

...

Keyword(s):

Cancer Cells ◽

Cancer Cell ◽

Immune Cell ◽

Human Cancer ◽

Antibody Fragment ◽

Cell Number ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Cell Therapies ◽

Human Cancer Cells

Chimeric antigen receptors (CARs) are synthetic receptors that reprogram T cells to kill cancer. The success of CAR-T cell therapies highlights the promise of programmed immunity and suggests that applying CAR strategies to other immune cell lineages may be beneficial. Here, we engineered a family of Chimeric Antigen Receptors for Phagocytosis (CAR-Ps) that direct macrophages to engulf specific targets, including cancer cells. CAR-Ps consist of an extracellular antibody fragment, which can be modified to direct CAR-P activity towards specific antigens. By screening a panel of engulfment receptor intracellular domains, we found that the cytosolic domains from Megf10 and FcRɣ robustly triggered engulfment independently of their native extracellular domain. We show that CAR-Ps drive specific engulfment of antigen-coated synthetic particles and whole human cancer cells. Addition of a tandem PI3K recruitment domain increased cancer cell engulfment. Finally, we show that CAR-P expressing murine macrophages reduce cancer cell number in co-culture by over 40%.

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The gene coding for secreted phosphoprotein1/osteopontin is the most overexpressed gene in cholangiocarcinoma—detected by oligonucleotide arrays and LightCycler analysis

Journal of Clinical Oncology ◽

10.1200/jco.2006.24.18_suppl.10036 ◽

2006 ◽

Vol 24 (18_suppl) ◽

pp. 10036-10036

Author(s):

H. G. Hass ◽

J. Jobst ◽

O. Nehls ◽

A. Frilling ◽

J. T. Hartmann ◽

...

Keyword(s):

Cell Function ◽

Immune Cell ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Cell Types ◽

Cell Phenotype ◽

Oligonucleotide Arrays ◽

Surface Receptors ◽

Malignant Liver ◽

And Control

10036 Background: Cholangiocarcinomas (CCC) are the second most common primary hepatic malignancy with a still poor prognosis and arise from biliary epithelia or cholangiocytes. Until now, less is known about the molecular pathways leeding to CCC. Methods: Oligonucleotide arrays were used to analyze gene expression profiles of 8 intrahepatic CCCs. After isolation of tRNA and transcription into cDNA, biotin-labelled cRNA probes were hybridized to GeneArrays (Affymetrix U 133A) containing probes of more than 22.000 genes/ESTs. For two-dimensional cluster analysis we used special software programs (Genexplore, GeneSpring). Dysregulated genes were determined by presence in more than 70% and a 2-fold change in relation to the corresponding non-malignant liver tissue. Lightcycler analysis were performed to validate the expression datas of dysregulated genes. Results: A total of 694 dysregulated genes (330 up-/364 down-regulated, compared with corresponding non-malignant tissue) were detected. As the gene with the highest and most consistent upregulation we were able to identify osteopontin (OPN) with an average 5-fold overexpression in all CCC tissues. OPN is an acidic phosphoprotein that is secreted by osteoblasts, macrophages and many other cell types and binds to a variety of cell surface receptors (integrins/CD44). OPN is multifunctional, with activities in cell migration, regulation of bone metabolism, immune cell function and control of tumor cell phenotype. Elevated OPN levels were seen in different tumors but until now no data exist about the expression in CCCs. As one possible interaction in human carcinogenesis, OPN has recently been shown to be a novel substrate for some MMPs, which play an importand role in tumor invasion and metastasis. Conclusions: This is the first report about an overexpression of OPN in CCC and our data indicate an important role in cholangiocarcinogenesis. Further studies are needed to illucidate the moleculargenetic mechanisms of OPN interactions in CCC. No significant financial relationships to disclose.

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Immune Escape Mechanisms and Future Prospects for Immunotherapy in Neuroblastoma

BioMed Research International ◽

10.1155/2018/1812535 ◽

2018 ◽

Vol 2018 ◽

pp. 1-11 ◽

Cited By ~ 20

Author(s):

Thitinee Vanichapol ◽

Somchai Chutipongtanate ◽

Usanarat Anurathapan ◽

Suradej Hongeng

Keyword(s):

Immune System ◽

T Cell ◽

Cell Therapy ◽

Cell Function ◽

Immune Cell ◽

Immune Escape ◽

Adoptive Cell Therapy ◽

Immune Recognition ◽

T Cell Therapy ◽

Cell Therapies

Neuroblastoma (NB) is the most common extracranial solid tumor in childhood with 5-year survival rate of 40% in high-risk patients despite intensive therapies. Recently, adoptive cell therapy, particularly chimeric antigen receptor (CAR) T cell therapy, represents a revolutionary treatment for hematological malignancies. However, there are challenges for this therapeutic strategy with solid tumors, as a result of the immunosuppressive nature of the tumor microenvironment (TME). Cancer cells have evolved multiple mechanisms to escape immune recognition or to modulate immune cell function. Several subtypes of immune cells that infiltrate tumors can foster tumor development, harbor immunosuppressive activity, and decrease an efficacy of adoptive cell therapies. Therefore, an understanding of the dual role of the immune system under the influences of the TME has been crucial for the development of effective therapeutic strategies against solid cancers. This review aims to depict key immune players and cellular pathways involved in the dynamic interplay between the TME and the immune system and also to address challenges and prospective development of adoptive T cell transfer for neuroblastoma.

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Fibroblasts – the neglected cell type in peripheral sensitization and chronic pain? - A systematic view on the current state of the literature

10.1101/2021.02.19.431978 ◽

2021 ◽

Author(s):

Naomi Shinotsuka ◽

Franziska Denk

Keyword(s):

Chronic Pain ◽

Cell Function ◽

Immune Cell ◽

Data Extraction ◽

Cell Types ◽

Extraction Methods ◽

Peripheral Sensitization ◽

Cell Type ◽

Systematic Screening ◽

Pain Research

AbstractChronic pain and its underlying biological mechanisms have been studied for many decades, with a myriad of molecules, receptors and cell types known to contribute to abnormal pain sensations. We now know that besides an obvious role for neuronal populations in the peripheral and central nervous system, immune cells like microglia, macrophages and T cells are also important drivers of persistent pain. While neuroinflammation has therefore been widely studied in pain research, there is one cell-type that appears to be rather neglected in this context: the humble fibroblast.Fibroblasts may seem unassuming, but actually play a major part in regulating immune cell function and driving chronic inflammation. What is known about them in the context chronic pain?Here we set out to analyze the literature on this topic – using systematic screening and data extraction methods to obtain a balanced view on what has been published. We found that there has been surprisingly little research in this area: 134 articles met our inclusion criteria, only a tiny minority of which directly investigated interactions between fibroblasts and peripheral neurons. We categorized the articles we included – stratifying them according to what was investigated, the estimated quality of results, and any common conclusions.Fibroblasts are a ubiquitous cell type and a prominent source of many pro-algesic mediators in a wide variety of tissues. We think that they deserve a more central role in pain research and propose a new, testable model of how fibroblasts might drive peripheral neuron sensitization.

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CARPOOL: A library-based platform to rapidly identify next generation chimeric antigen receptors

10.1101/2021.07.09.450900 ◽

2021 ◽

Author(s):

Taeyoon Kyung ◽

Khloe S Gordon ◽

Caleb R Perez ◽

Patrick V Holec ◽

Azucena Ramos ◽

...

Keyword(s):

High Throughput Screening ◽

Cell Types ◽

Tumor Control ◽

Antigen Receptors ◽

Cell Therapies ◽

Promising Tool ◽

Signaling Domain ◽

Screening Platform

CD19-targeted CAR therapies have successfully treated B cell leukemias and lymphomas, but many responders later relapse or experience toxicities. CAR intracellular domains (ICDs) are key to converting antigen recognition into anti-tumor effector functions. Despite the many possible immune signaling domain combinations that could be included in CARs, almost all CARs currently rely upon CD3𝛇, CD28, and/or 4-1BB signaling. To explore the signaling potential of CAR ICDs, we generated a library of 700,000 CD19 CAR molecules with diverse signaling domains and developed a high throughput screening platform to enable optimization of CAR signaling for anti-tumor functions. Our strategy identifies CARs with novel signaling domain combinations that elicit distinct T cell behaviors from a clinically available CAR, including enhanced proliferation and persistence, lower exhaustion, potent cytotoxicity in an in vitro tumor rechallenge condition, and comparable tumor control in vivo. This approach is readily adaptable to numerous disease models, cell types, and selection conditions, making it a promising tool for rapidly improving adoptive cell therapies and expanding their utility to new disease indications.

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CAR-T Therapy for Solid Tumors: Development of New Strategies

Trends in Immunotherapy ◽

10.24294/ti.v4.i2.1064 ◽

2020 ◽

Vol 4 (2) ◽

pp. 41

Author(s):

Marvin De los Santos ◽

Samuel D. Bernal

Keyword(s):

Solid Tumors ◽

New Technologies ◽

Immune Cell ◽

Significant Development ◽

Cell Therapies ◽

Car T Cells ◽

Car T ◽

Cancer Types ◽

Enhance Efficiency ◽

New Strategies

The recent approval of two CAR-T therapies by US Food and Drug Administration (FDA) marks a very significant development in cell-based cancer immunotherapy. This milestone was demonstrated by the effectiveness of eradicating hematologic cancers using CD19-specific CARs. The success spurred development of immune cell therapies for other cancers, especially solid tumors. The generation of novel CAR constructs for these cancer types represents a major challenge in bringing the technology ‘from-bench-to-bedside‘.In this review, we outline some new technologies we have developed to equip CAR-T cells to enhance efficiency while decreasing toxicity of CAR-T therapies in solid tumors.

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An Alternative Cell Therapy for Cancers: Induced Pluripotent Stem Cell (iPSC)-Derived Natural Killer Cells

Biomedicines ◽

10.3390/biomedicines9101323 ◽

2021 ◽

Vol 9 (10) ◽

pp. 1323

Author(s):

Li-Jie Hsu ◽

Chao-Lin Liu ◽

Ming-Ling Kuo ◽

Chia-Ning Shen ◽

Chia-Rui Shen

Keyword(s):

Cell Therapy ◽

Nk Cells ◽

Natural Killer ◽

Immune Cell ◽

Ex Vivo ◽

Induced Pluripotent Stem Cell ◽

Cell Types ◽

The Body ◽

Cell Therapies ◽

Induced Pluripotent

Cell therapy is usually defined as the treatment or prevention of human disease by supplementation with cells that have been selected, manipulated, and pharmacologically treated or altered outside the body (ex vivo). Induced pluripotent stem cells (iPSCs), with their unique characteristics of indefinite expansion in cultures and genetic modifications, represent an ideal cell source for differentiation into specialized cell types. Cell therapy has recently become one of the most promising therapeutic approaches for cancers, and different immune cell types are selected as therapeutic platforms. Natural killer (NK) cells are shown to be effective tumor cell killers and do not cause graft-vs-host disease (GVHD), making them excellent candidates for, and facilitating the development of, “off-the-shelf” cell therapies. In this review, we summarize the progress in the past decade in the advent of iPSC technology and review recent developments in gene-modified iPSC-NK cells as readily available “off-the-shelf” cellular therapies.

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Reflections on Thyroid Autoimmunity: A Personal Overview from the Past into the Future

Hormone and Metabolic Research ◽

10.1055/a-0725-9297 ◽

2018 ◽

Vol 50 (12) ◽

pp. 840-852 ◽

Cited By ~ 7

Author(s):

Basil Rapoport ◽

Sandra McLachlan

Keyword(s):

Antigen Processing ◽

Plasma Cells ◽

Immune Cell ◽

Thyroid Autoimmunity ◽

Cell Types ◽

Autoimmune Response ◽

Personal Perspective ◽

Graves Disease ◽

Antigen Receptors ◽

Thyroid Autoantigens

AbstractAfter investigating thyroid autoimmunity for more than 40 years, we present a personal perspective on the field. Despite effective therapies for Graves’ hyperthyroidism and Hashimoto’s thyroiditis, cures are elusive. Novel forms of therapy are being developed, such as small molecule inhibitors of the TSH receptor (TSHR), but cure will require immunotherapy. This goal requires advances in understanding the pathogenesis of thyroid autoimmunity, the ‘keys’ for which are the thyroid antigens themselves. Presently, however, greater investigative focus is on non-thyroid specific immune cell types and molecules. Thyroid autoantigens are the drivers of the autoimmune response, a prime example being the TSHR. In our view, the TSHR is the culprit as well as the victim in Graves’ disease because of its unique structure. Unlike the closely related gonadotropin receptors, the TSHR cleaves into subunits and there is strong evidence that its shed extracellular A-subunit, not the holoreceptor, is the major antigen driving pathogenic thyroid stimulating autoantibodies (TSAb) development. There is no Graves’ disease of the gonads. Studies of potential antigen-specific immunotherapies require an animal model. Such models have been developed in which TSAb can be induced or, more importantly, arise spontaneously. Not appreciated until recently by thyroid investigators is that B cell surface autoantibodies are highly efficient ‘antigen receptors’ and the epitope to which an autoantibody binds influences antigen processing and which peptide is presented to T cells. These animal models and recombinant human autoantibodies cloned from Graves’ and Hashimoto’s B cells (plasma cells) are available for study by future generations.

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Nanoengineering of Immune Cell Function

MRS Proceedings ◽

10.1557/proc-1209-yy03-01 ◽

2009 ◽

Vol 1209 ◽

Cited By ~ 3

Author(s):

Keyue Shen ◽

Michael C Milone ◽

Michael L. Dustin ◽

Lance Cameron Kam

Keyword(s):

T Cells ◽

Immune System ◽

T Cell ◽

T Cell Activation ◽

Cell Activation ◽

Cell Function ◽

Immune Cell ◽

Cell Communication ◽

Cell Therapies ◽

Nano Scale

AbstractT lymphocytes are a key regulatory component of the adaptive immune system. Understanding how the micro- and nano-scale details of the extracellular environment influence T cell activation may have wide impact on the use of T cells for therapeutic purposes. In this article, we examine how the micro- and nano-scale presentation of ligands to cell surface receptors, including microscale organization and nanoscale mobility, influences the activation of T cells. We extend these studies to include the role of cell-generated forces, and the rigidity of the microenvironment, on T cell activation. These approaches enable delivery of defined signals to T cells, a step toward understanding the cell-cell communication in the immune system, and developing micro/nano- and material- engineered systems for tailoring immune responses for adoptive T cell therapies.

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