MCAM(CD146) Chimeric Antigen Receptor (CAR): A Comprehensive Guide and Our Service & Product Introduction

MCAM (Melanoma Cell Adhesion Molecule, CD146), also known as MUC18 or Gicerin, is a key cell adhesion molecule and tumor-associated antigen (TAA) that plays critical roles in angiogenesis, tumor progression, and immune regulation. As a professional provider of gene engineering tools, our company offers a comprehensive range of high-quality MCAM (CD146) CAR expression plasmid vectors and personalized custom construction services. We are dedicated to supporting researchers, biopharmaceutical enterprises, and clinical institutions in accelerating the development of MCAM (CD146)-targeted immunotherapies for various malignancies and diseases. If you are interested in our MCAM (CD146) CAR expression plasmid vectors or custom construction services, or need technical advice for your MCAM (CD146) CAR research, please contact us at admin@rgbiotech.com. Our professional team will promptly respond to your inquiries, providing personalized solutions to meet your research needs. We look forward to cooperating with you to advance MCAM (CD146) CAR technology and accelerate the development of innovative immunotherapies for MCAM-related solid tumors and angiogenesis-related diseases.

Our MCAM(CD146) CAR Expression Plasmid Vector Products and Custom Services

MCAM (CD146) CAR technology is a rapidly growing field in solid tumor immunotherapy, with increasing demand for high-quality plasmid vectors and custom construction services. As a leading provider of gene engineering tools, RGBiotech offers a comprehensive range of MCAM (CD146) CAR expression plasmid vectors and professional custom construction services, tailored to meet the diverse needs of researchers and biopharmaceutical companies. Our products and services are designed to accelerate MCAM (CD146) CAR research and clinical translation, with strict quality control and full-cycle technical support.

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Custom MCAM (CD146) CAR Plasmid Vector Construction Services

Introduction of MCAM(CD146)

The MCAM gene (official symbol: MCAM; HGNC ID: 7114; Gene ID: 4162; also known as CD146, Gicerin, MUC18, HEMCAM, METCAM) is located on human chromosome 11q23.3, spanning approximately 25 kb of genomic DNA. It belongs to the immunoglobulin superfamily of cell adhesion molecules, with a high degree of evolutionary conservation across mammalian species. The human MCAM gene has a reference sequence of NM_006500.2, with a gene length of 1941 bp, encoding a 113 kDa transmembrane glycoprotein composed of 646 amino acids. Multiple alternatively spliced transcript variants have been identified, producing two main isoforms (MCAM-L long and MCAM-S short) that differ in the length of their cytoplasmic domains and exhibit distinct functional properties. The gene’s 5’-flanking region contains conserved transcription regulatory elements that control its tissue-specific expression, with abnormal upregulation frequently observed in malignant tumors.

MCAM (CD146) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily, with a structure specifically adapted for cell adhesion and signal transduction. Its structure consists of five extracellular immunoglobulin-like (Ig) domains (one V-type and four C2-type), a single transmembrane domain, and a cytoplasmic tail that varies between the two isoforms. The extracellular Ig domains are responsible for ligand binding (e.g., laminin alpha 4) and cell-cell adhesion, while the transmembrane domain anchors the protein to the cell membrane. The long isoform (MCAM-L) has a cytoplasmic domain containing two protein kinase C (PKC) phosphorylation sites and a dileucine motif, while the short isoform (MCAM-S) contains one PKC phosphorylation site and a PDZ-binding domain, leading to functional differences in cell adhesion and signaling. A soluble form of MCAM (sCD146) is also produced by shedding of the extracellular portion via matrix metalloproteinases (MMPs), which plays a role in pathological angiogenesis and inflammation.

MCAM (CD146) primarily functions as a cell adhesion molecule, mediating homotypic (MCAM-MCAM) and heterotypic (MCAM-laminin alpha 4, MCAM-VEGFR-2) cell-cell and cell-matrix interactions. In normal physiology, it plays a critical role in vascular homeostasis, angiogenesis, embryonic development, wound healing, and immune regulation. Specifically, it is involved in endothelial cell junction formation, smooth muscle cell recruitment, and the regulation of mesenchymal stem cell multipotency, with higher MCAM expression correlating with greater differentiation potential in mesenchymal stem cells. MCAM is also expressed on a small subset of effector memory T cells that produce IL-17, contributing to the inflammatory response. In pathological conditions, abnormal MCAM overexpression promotes angiogenesis, tumor cell invasion, metastasis, and immune suppression, making it a key target for therapeutic intervention.

In normal adult tissues, MCAM (CD146) expression is primarily restricted to cells of the vascular system, including endothelial cells (all vessel sizes and anatomic locations), smooth muscle cells, and pericytes, where it maintains vascular architecture and function. It is also expressed at low levels on a small subset of peripheral blood mononuclear cells (≈1%), including T cells, B cells, and NK cells, as well as on mesenchymal stem cells, trophoblasts, and activated T cells. In contrast, MCAM is widely overexpressed in various malignant tumors, with high expression levels correlating with disease progression and metastasis. Notably, it is overexpressed in melanoma, osteosarcoma, synovial sarcoma, Kaposi’s sarcoma, neuroblastoma, breast cancer, lung cancer, and colorectal cancer, making it an ideal target for targeted immunotherapy.

MCAM (CD146) is closely associated with the development, progression, and prognosis of various tumors and non-malignant diseases, primarily due to its role in angiogenesis and cell adhesion.
1) Malignant Tumors: MCAM overexpression is a hallmark of multiple solid tumors, including melanoma (where it was first identified), osteosarcoma, synovial sarcoma, Kaposi’s sarcoma, neuroblastoma, breast cancer, non-small cell lung cancer (NSCLC), and colorectal cancer. In these tumors, MCAM promotes tumor angiogenesis, invasion, and metastasis, and its high expression correlates with poor prognosis and resistance to conventional therapies. 2) Angiogenic and Inflammatory Diseases: Abnormal MCAM expression is involved in pathological angiogenesis associated with autoimmune diseases, obstetric and ocular pathologies, cardiovascular disorders, and chronic inflammatory diseases (e.g., inflammatory bowel disease, chronic obstructive pulmonary disease). Soluble CD146 (sCD146) levels are significantly elevated in the serum or interstitial fluid of patients with these conditions, contributing to disease progression. 3) Vascular Disorders: MCAM dysregulation is linked to vascular dysfunction, including atherosclerosis and restenosis, due to its role in endothelial cell junction stability and smooth muscle cell proliferation.

MCAM (CD146) Chimeric Antigen Receptor (CAR) is a genetically engineered receptor designed to redirect immune cells (e.g., T cells, NK cells, macrophages) to specifically recognize and eliminate MCAM (CD146)-expressing tumor cells and abnormal vascular cells. By fusing the extracellular MCAM-binding domain (e.g., anti-MCAM single-chain variable fragment (scFv)) with intracellular signaling domains, MCAM (CD146) CAR-equipped immune cells achieve MHC-independent recognition of MCAM-positive cells, overcoming the immunosuppressive tumor microenvironment and providing a promising strategy for treating MCAM-positive tumors and angiogenesis-related diseases.

Introduction of MCAM(CD146) Chimeric Antigen Receptor (CAR)

A typical MCAM (CD146) CAR consists of three core components, optimized for targeting MCAM-positive tumors and vascular cells.
1) Extracellular Antigen-Binding Domain: Usually a high-affinity anti-MCAM (CD146) scFv or MCAM ligand (e.g., laminin alpha 4 fragment), responsible for specific binding to the extracellular Ig domains of MCAM on target cells. Fully human scFv domains are preferred to reduce immunogenicity and improve in vivo persistence.
2) Hinge and Transmembrane Domain: Derived from CD28 or CD8α, facilitating the stability of the CAR on the immune cell surface, reducing steric hindrance in the tumor microenvironment, and enabling efficient transmission of intracellular signals.
3) Intracellular Signaling Domain: Varies by CAR generation, including activation domains (CD3ζ) and co-stimulatory domains (CD28, 4-1BB, OX40, etc.), which regulate the activation, proliferation, persistence, and anti-tumor activity of CAR-engineered cells. Advanced designs may include cytokine fusions or immune checkpoint antagonist domains to enhance efficacy in solid tumors and overcome T cell exhaustion.

Current Research Achievements of MCAM (CD146) CAR

Approved Drugs of MCAM (CD146) CAR

MCAM (CD146) CAR Research Hotspots

MCAM (CD146) CAR Research Difficulties & Challenges

References

Q: What is the difference between different generations of MCAM (CD146) CARs, and how to choose the right one?
A: The main difference lies in the intracellular signaling domains. 1st generation CARs (CD3ζ only) are suitable for preliminary studies of MCAM recognition but have low in vivo persistence and are prone to exhaustion. 2nd (CD3ζ + single co-stimulatory domain) and 3rd (CD3ζ + two co-stimulatory domains) generations are ideal for preclinical studies requiring enhanced cell activation and persistence, especially for in vivo models. 4th generation CARs (with cytokines) are suitable for solid tumor research to improve tumor infiltration and persistence, addressing the TME barrier. 5th generation CARs (with checkpoint antagonists) are designed to overcome T cell exhaustion, making them ideal for long-term disease control in advanced solid tumors. Choose based on your research goals (e.g., melanoma vs. osteosarcoma, in vitro vs. in vivo studies) and cell type (T cells vs. NK cells vs. macrophages).

Q: Which vector backbone is suitable for MCAM (CD146) CAR-T/NK cell preparation, and why?
A: Lentiviral vectors are the most commonly used backbone for MCAM (CD146) CAR-T/NK cell preparation, as they can transduce both dividing and non-dividing cells, integrate into the host genome for stable long-term CAR expression, and have low immunogenicity-critical for avoiding human anti-mouse antibody (HAMA) responses. Retroviral vectors are suitable for dividing cells (e.g., activated T cells) and are cost-effective for large-scale production. AAV vectors are ideal for transient or tissue-specific MCAM CAR expression, with high safety and low integration risk. Non-viral plasmid vectors are suitable for in vitro studies or non-viral delivery systems (e.g., electroporation, LNP), reducing the risk of viral vector-related complications and lowering manufacturing costs-particularly for off-the-shelf therapies.

Q: How to ensure high MCAM (CD146) CAR expression efficiency in target cells?
A: Several factors affect expression efficiency: 1) Choose a suitable promoter (EF-1α for stable long-term expression, CMV for strong transient expression, or tumor-specific promoters for targeted expression); 2) Optimize the MCAM (CD146) CAR gene codons for the target cell type (e.g., human primary T cells) to enhance translation efficiency; 3) Use high-purity, low-endotoxin plasmids to avoid cytotoxicity that impairs cell viability and expression; 4) Select an appropriate delivery method; 5) Ensure the vector has a suitable linker (2A or IRES) for co-expression of CAR and markers, avoiding interference between components.

Q: What are the key considerations for MCAM (CD146) CAR plasmid vector storage and handling?
A: To maintain plasmid integrity and activity: 1) Store plasmids at -20°C or -80°C, avoiding repeated freeze-thaw cycles (aliquot into single-use portions to prevent degradation); 2) Thaw plasmids on ice and centrifuge briefly before use to ensure uniform concentration; 3) Avoid exposure to high temperatures, UV light, and RNase/DNase contamination, which can damage plasmid DNA; 4) For lentiviral/retroviral backbones, store the packaged virus at -80°C with cryoprotectants to maintain transduction efficiency; 5) Follow sterile procedures when handling plasmids for cell culture applications to prevent contamination that could invalidate experiments; 6) Lyophilized plasmids can be stored at ambient temperature for three months, facilitating transportation and short-term storage.

Q: Can your custom service design MCAM (CD146) CAR vectors with specific modifications (e.g., isoform-specific targeting, bispecific design, cytokine fusion)?
A: Yes. Our custom service supports various modifications, including designing isoform-specific CARs to target MCAM-L (the tumor-associated isoform), fusing cytokines (IL-7, IL-15, IL-21) to improve in vivo expansion and persistence, and designing bispecific CARs (e.g., MCAM/CD44v6) to mitigate tumor escape. Simply provide your modification requirements, and our team will design a tailored solution.

Q: How to verify the functionality of MCAM (CD146) CAR plasmid vectors after purchase?
A: We recommend the following verification steps: 1) Sequence verification to confirm the plasmid sequence is correct, including the MCAM (CD146) CAR gene, promoter, and markers; 2) Transfect HEK293T cells with the plasmid, then detect MCAM (CD146) CAR expression via flow cytometry (using anti-MCAM or anti-CAR antibodies) or fluorescence microscopy (if fluorescent markers are included)-similar to the validation methods used for MCAM expression vectors; 3) Perform in vitro cytotoxicity assays using MCAM-expressing target cells (e.g., melanoma, osteosarcoma, neuroblastoma cell lines) to verify the killing ability of CAR-transduced T/NK cells; 4) Detect cytokine production (e.g., IFN-γ, IL-2) via ELISA to confirm CAR activation.

Q: What is the difference between MCAM (CD146) CAR-T and MCAM (CD146) CAR-NK cells, and which is better for my research?
A: MCAM (CD146) CAR-T cells have strong proliferation ability and long-term persistence, making them suitable for long-term disease control (e.g., advanced melanoma, osteosarcoma) but may have higher risk of CRS and difficulty infiltrating solid tumors. MCAM (CD146) CAR-NK cells have lower immunogenicity, no risk of graft-versus-host disease (GVHD), and rapid cytotoxicity, making them suitable for patients with compromised T cell function or acute advanced disease. They also have better penetration into solid tumors and lower CRS risk, with enhanced efficacy when combined with IL-15 agonists such as NKTR-255. However, CAR-NK cells have shorter in vivo persistence and may require repeated infusions. Choose based on your research focus (long-term control vs. rapid efficacy) and safety requirements (e.g., allogeneic vs. autologous therapy).

Q: How to reduce off-target effects of MCAM (CD146) CAR-engineered cells?
A: Off-target effects can be reduced by: 1) Using highly specific extracellular binding domains (e.g., fully human anti-MCAM scFv) that only recognize MCAM on tumor cells or the MCAM-L isoform; 2) Using tumor-specific or vascular cell-specific promoters to restrict CAR expression to target cells, avoiding expression in normal MCAM-positive vascular cells; 3) Adding a suicide gene (e.g., iCasp9) to the vector to eliminate CAR-engineered cells if off-target toxicity occurs; 4) Designing bispecific CARs that target MCAM and a tumor-specific antigen, enhancing specificity for malignant cells; 5) Optimizing CAR affinity to avoid cross-reactivity with other immunoglobulin superfamily members.

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