All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.
Chimeric antigen receptors (CARs) represent a groundbreaking advancement in the fight against cancer and other diseases. Engineered receptors endow T cells with the ability to specifically target and eliminate malignant cells, offering a promising avenue for personalized and efficient therapies. Researchers face numerous challenges in developing CARs that are highly specific and effective, while minimizing potential off-target effects. Each step, from selecting appropriate antigen-binding domains to optimizing CAR structural components, is crucial for creating therapeutic agents that achieve expected clinical outcomes.
Understanding these complexities, we are committed to providing comprehensive CAR design and construction services tailored to meet the unique requirements of each project. Whether it's conventional CAR designs from first-generation to fifth-generation CARs, or novel designs like TanCARs, Physiological CARs, and Universal CARs, our experienced team employs innovative thinking to ensure each CAR we help design maximizes efficacy and safety.
Utilizing advanced bioinformatics tools and molecular biology techniques, we guide clients through the complex CAR development process from initial conceptualization to final validation. Our services are designed to accelerate research and development timelines, facilitating swift transitions from the lab to clinical applications. Whether your focus is on hematologic malignancies, solid tumors, or exploring CAR applications in autoimmune diseases, our goal is to support your work by providing cutting-edge solutions and expert guidance. Partner with us to access rich knowledge and resources dedicated to advancing the frontier of cellular immunotherapy.
The original CAR-T or the first-generation CAR consists of the intracellular domain from the CD3 ζ- chain and the primary transmitter of signals from endogenous TCRs, which showed success in pre-clinical trials and entered Phase I clinical trials in ovarian cancer, neuroblastoma and various types of leukemia and lymphoma. Although the anti-tumor activity was limited due to insufficient activation, persistence and homing to the cancer tissue, some significant effects indeed existed in patients with B-cell lymphoma treated with α-CD20-CD3 ζ CAR-modified T cells and some neuroblastoma treated with ScFv-CD3 ζ CAR-Ts. This is the most common form of CARs to fuse single-chain variable fragments (scFv) derived from monoclonal antibodies to CD3 ζ transmembrane and endo domain.
To augment the antitumor efficacy of 1st-generation CARs, the 2nd-generation CARs were designed to combine the intracellular signaling domains from various costimulatory protein receptors (e.g., CD28, 41BB, ICOS) incorporated in the cytoplasmic tail of the CAR to enhance the signaling. For instance, the CD19-targeted CARs incorporated with CD28 or 4-1BB signaling domains manifested remarkable complete remission rates in patients with refractory B-cell malignancies. Subsequently, the CD28-based CARs showed a brisk proliferative response and boost effector functions. Meanwhile, the 4-1BB-based CARs manifested a more progressive T cell accumulation.
As the expectation of more antitumor efficacy, the 3rd-generation of CARs combined multiple signaling domains (e.g., CD3 ζ-CD28-41BB, CD3 ζ-CD28-OX40) to acquire further enhanced activation signals, proliferation, production of cytokines and effective function. For instance, the α-CD19-CD3 ζ-4-1BB CAR-Ts for chronic lymphocyte leukemia showed complete remission to infiltrate and lyse cancer tissue. Even better, a fraction of CAR-Ts functioned as a memory phenotype for preventing tumor relapses. Despite the significant therapeutic effect, the emerging uncontrollable activity accompanied with more antitumor efficacy caused life-threatening lysis activity as the most critical adverse effect or toxicity including clinically significant release of pro-inflammatory cytokines, pulmonary toxicity, multi-organ failure, and eventual death.
The previous CAR strategies are highly specific and useful in redirecting T cells targeting malicious cancer cells. However, the major limitation on solid tumors with a tremendous phenotypic heterogeneity and relapse due to antigen-negative cancer cells is the huge challenge to trigger a novel CAR strategy. The 4th-generation CAR is designed to shape the tumor environment by the inducible release of transgenic immune modifiers, such as IL-12, which augments T-cell activation, attracts and activates innate immune cells to eliminate antigen-negative cancer cells in the targeted lesion.
The fifth generation of CARs, currently under exploration, builds upon the second generation but includes a truncated cytoplasmic interleukin-2 (IL-2) receptor β-chain domain (IL-2Rβ) with a binding site for the signal transducer and activator of transcription-3 (STAT3). The antigen-specific activation of this receptor triggers three critical signaling pathways simultaneously: TCR signaling through the CD3ζ domains, co-stimulatory signaling via the CD28 domain, and cytokine signaling through the JAK–STAT3/5 pathway. This integrated approach provides the synergistic signals necessary for full T cell activation and proliferation, effectively enhancing the immune response.
Dual CAR T cells improve the specificity and safety of CAR-T therapy by targeting two tumor-associated antigens simultaneously. They are designed to activate only when both antigens are present on the tumor, reducing off-tumor toxicity. These cells express two CARs: one binding to a specific tumor antigen and delivering the primary signal, and another binding to a different antigen and providing the co-stimulatory signal. For example, dual CAR-T cells co-expressing anti-ErbB2 and anti-MUC1 CARs use CD3ζ and CD28 signaling pathways, respectively, and effectively kill ErbB2+ tumor cells only in the presence of MUC1. This approach enhances therapeutic efficacy while minimizing effects on normal tissues expressing a single antigen.
Conditional CAR-T cells enhance the safety of CAR-T cell therapy by requiring an additional signal for activation beyond target recognition. This secondary signal can be a small molecule or an adaptor-specific receptor that binds to secondary antibodies directed at target cell antigens. Conditional CARs act as a switch, activated by an exogenous molecule, allowing precise regulation of CAR-T cell activity without leading to cell death in its absence. For example, a conditional CAR targeting CD19 in a Nalm-6 xenograft rodent model demonstrated controlled activity, tissue-homing, cytokine release, and phenotype in a dose-titratable manner, potentially reducing cytokine release syndrome compared to conventional CAR-T cells.
TanCARs are an advanced type of chimeric antigen receptor designed to enhance the efficacy of CAR-T cell therapy. Unlike traditional CARs, TanCARs feature two distinct antigen recognition domains on a single transgenic receptor. This allows for simultaneous targeting of tumor cells and elements within the tumor microenvironment, increasing T cell activation and function by enhancing avidity and broadening therapeutic reach. This dual targeting capability addresses issues like antigen-loss escape variants commonly seen in cancer cells, potentially leading to more effective and durable cancer treatments.
Unlike traditional CARs, which often use single-chain variable fragments (scFv) of murine origin and risk triggering immune responses, physiological CARs utilize receptor ligands such as HER3 and HER4. These CARs consist of an antigen receptor and a CD3ζ intracellular signaling domain, with or without a transmembrane and spacer region, engineered into immune cells to target ligands expressed on tumor cells. This approach enhances the ability of T cells to recognize and eliminate cancer cells, increasing the persistence and effectiveness of the therapy while reducing the risk of immune response and anaphylaxis associated with murine-based scFv CARs.
Universal CARs represent a significant advancement in CAR-T cell therapy by offering flexibility and adaptability to target a wide range of malignancies. These CARs are engineered to recognize various tagged antigens, such as biotin-labeled molecules, FITC-labeled targets, and peptide nucleic acids, among others. This versatility allows for precise and personalized treatment options, enhancing T cell activation, controlling therapeutic effects, and reducing adverse reactions. Universal CARs improve the scope and efficacy of CAR-T therapies, making them more effective against diverse cancer types.
Marked CAR-T cells are specially designed CAR-T cells that express both CARs and tumor epitopes that are recognized by existing monoclonal antibodies. This dual expression allows for a unique safety mechanism: if severe adverse effects occur, the specific monoclonal antibody can be administered to selectively clear the CAR-T cells, thereby avoiding additional off-tumor effects. This approach not only helps in mitigating side effects but also enhances the overall therapeutic potential of CAR-T cell treatments by providing a controlled and targeted method to manage and eliminate CAR-T cells if necessary. In essence, Marked CARs combines efficacy with improved safety, offering a promising solution for cancer treatment.
For more CAR design and construction, please visit our Smart™ CAR Construction Service:
Background
Solid tumors present significant challenges to immunotherapy due to their immunosuppressive microenvironment. Conventional CAR-T cell therapies have shown limited efficacy against solid malignancies, necessitating novel approaches to enhance their effectiveness. The study aimed to develop a method to remodel the tumor microenvironment, enabling CAR-T cells to effectively target and destroy solid tumors.
Methodology
The researchers utilized a murine 4T1 breast cancer cell line and a glioma mouse model to study the effects of combining targeted lipid nanoparticles with CAR-T cell therapy. The nanoparticles were designed to deliver a combination of a PI3K inhibitor (PI-3065) to inhibit immune-suppressive tumor cells and an NKT cell agonist (7DW8-5) to stimulate therapeutic T cells. The nanoparticles were administered in repeated infusions and were targeted to the tumor site using the iRGD peptide. Following nanoparticle preconditioning, tumor-specific CAR-T cells were infused during a therapeutic window created by the preconditioning. The effectiveness of the treatment was measured by assessing tumor regression, T-cell infiltration, and survival rates in treated animals.
Results
Conclusion
The study demonstrated that the use of targeted lipid nanoparticles to precondition the tumor microenvironment significantly enhances the efficacy of CAR-T cell therapy against solid tumors. This novel approach overcomes the traditional barriers posed by the immunosuppressive milieu of solid tumors, leading to improved T-cell infiltration, robust antitumor activity, and extended survival. The findings suggest that nanoparticle-mediated preconditioning could be a practical and low-cost strategy to potentiate various cancer immunotherapies, including CAR-T cell therapy, vaccines, and bispecific T-cell engager (BITE) platforms. This research offers a promising avenue for improving clinical outcomes in patients with solid malignancies.
Creative Biolabs' Contribution
Creative Biolabs provided custom-designed chimeric antigen receptor (CAR) constructs essential for this study. The anti-ROR1-28z CAR and the anti-EGFRvIII-28z CAR constructs were tailored to target tyrosine kinase-like orphan receptor 1 (ROR1) and epidermal growth factor receptor variant III (EGFRvIII), respectively. These CAR constructs consisted of:
These sophisticated CAR constructs were crucial for the genetic modification of T cells, enabling them to effectively target and attack tumor cells expressing ROR1 and EGFRvIII. Creative Biolabs' expertise in designing and producing these high-affinity CAR constructs provided the foundation for the study's success, showcasing the pivotal role of advanced biotechnology services in overcoming the barriers posed by solid tumor immunotherapy.
Use the resources in our library to help you understand your options and make critical decisions for your study.
Chimeric antigen receptor (CAR) can combine the extracellular antigen recognition domain from antibodies with the immune cell signaling domain to redirect T cell specificity and induce potent antitumor activity. CAR is an artificial transmembrane receptor that connects the extracellular antigen recognition domain, hinge domain (HD), transmembrane domain (TMD), and intracellular signal transduction domain in series.
CAR-T cell therapies revolutionize cancer treatment by harnessing the power of a patient's immune cells. Engineered with chimeric antigen receptors (CARs), these cells effectively target and destroy cancer cells, offering a personalized and potent approach. This groundbreaking immunotherapy has shown remarkable success in treating certain blood cancers, providing hope for patients who may not respond to traditional treatments. CAR-T cell therapies mark a significant stride towards precision medicine, ushering in a new era in oncology with the potential to transform the landscape of cancer care.
Based on the significant roles of T cells in the immune system, many small-molecule drugs targeting T cells and T-cell based immunotherapies have been developed for the treatment of intractable diseases including autoimmune diseases and cancer. T cell-based immunotherapies mainly utilize the mechanisms of T cell-mediated immune responses and the effects of some other immune cells such as dendritic cells (DCs), natural killer (NK) cells, and macrophages.
Adoptive cell transfer (ACT) of engineered T cells is a cutting-edge therapeutic approach revolutionizing cancer treatment. This innovative method involves modifying T cells, a key component of the immune system, to enhance their ability to target and eliminate cancer cells. By introducing genetically engineered T cells into patients, researchers aim to bolster the immune response against cancer, offering a personalized and potentially curative treatment option. This groundbreaking technology holds promise for addressing various malignancies and represents a significant stride towards more effective and precise cancer therapies.
Aiming at clearing the tumor without the toxicity of conventional treatments, Creative Biolabs inherits advantages of CAR-T strategy to help researchers make scientific history fighting malicious cancers.
For any technical issues or product/service related questions, please leave your information below. Our team will contact you soon.
The latest newsletter to introduce the latest breaking information, our site updates, field and other scientific news, important events, and insights from industry leaders
LEARN MORE NEWSLETTERCellRapeutics™ In Vivo Cell Engineering: One-stop in vivo T/B/NK cell and macrophage engineering services covering vectors construction to function verification.
LEARN MORE SOLUTIONSilence™ CAR-T Cell: A novel platform to enhance CAR-T cell immunotherapy by combining RNAi technology to suppress genes that may impede CAR functionality.
LEARN MORE NOVEL TECHNOLOGYCanine CAR-T Therapy Development: From early target discovery, CAR design and construction, cell culture, and transfection, to in vitro and in vivo function validation.
LEARN MORE SOLUTION