CellRapeutics™ CAR-Treg Development Service

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Chimeric antigen receptor-modified regulatory T cells (CAR-Tregs) represent a groundbreaking frontier in cellular immunotherapy, combining the specificity of CAR technology with the powerful immunosuppressive capabilities of regulatory T cells. This innovative therapeutic approach holds immense promise for treating autoimmune diseases, preventing transplant rejection, and managing various inflammatory conditions. By engineering Tregs to express CARs specific to desired antigens, these cells can provide targeted immunosuppression while maintaining immune homeostasis in specific tissues, offering superior therapeutic potential compared to conventional treatments.

At Creative Biolabs, we leverage our extensive expertise in CAR technology and immunology to offer comprehensive CAR-Treg development services. Our end-to-end solutions, backed by state-of-the-art facilities and experienced scientists, enable partners to create CAR-Tregs products that support the next generation of cell therapy innovation.

Fig. 1 The structure of CAR-Tregs and their suppression of effector T cells. Fig. 1 Schematic diagram depicting the structure of CAR-Tregs and their suppression of effector T (Teff) cells1.

CAR-T Cells vs CAR-Treg Cells

The distinct functional variations between conventional CAR-T cells and CAR-Treg cells present unique opportunities for treating different diseases. Below, we examine the fundamental differences between these therapeutic approaches, highlighting their distinct characteristics, challenges, and advantages.

Conventional CAR-T Cells

Cell Origin & Properties

Derived from effector T cells (CD8+ cytotoxic T cells), engineered to recognize and eliminate specific target cells through direct cytotoxicity.
Mechanism of Action

Direct cytotoxicity through perforin and granzyme release, accompanied by pro-inflammatory cytokine production (IFN-γ, TNF-α), leading to potential cytokine release syndrome (CRS).
Clinical Applications

Primarily successful in treating hematological malignancies such as refractory acute lymphoblastic leukemia and relapsed/refractory non-Hodgkin's lymphoma, with ongoing clinical trials exploring applications in solid tumors.

Key Challenges

Risk of severe cytokine release syndrome (CRS) with elevated inflammatory markers and multi-organ dysfunction.
Limited to small clinical trials due to potentially fatal side effects including neurotoxicity and systemic inflammation.
Possible transformation into immunosuppressive inducible Treg cells during tumor metastasis, compromising therapeutic efficacy.
Potential for on-target, off-tumor effects leading to unintended tissue damage and organ dysfunction.

CAR-Treg Cells

Cell Origin & Properties

Engineered from regulatory T cells (CD4⁺ CD25⁺ FOXP3⁺), specifically designed for maintaining immune tolerance and homeostasis.
Mechanism of Action

Targeted immunosuppression through anti-inflammatory cytokines (IL-10, TGF-β), with direct suppression of effector T cells and modulation of antigen-presenting cells.
Clinical Applications

Specifically designed for autoimmune diseases, transplant rejection prevention, and inflammatory conditions, offering targeted immunosuppression.

Key Advantages

Superior antigen-specific inhibition compared to polyclonal Tregs
Stable phenotype with targeted migration to disease-associated organs
Reduced side effects due to antigen-specific immunosuppression
MHC-independent function with reduced IL-2 dependency compared to TCR-Treg
Avoids systemic immunosuppression associated with polyclonal Tregs

Our Services

From initial biomarker identification to clinical trial support, we offer a highly customizable service workflow tailored to your specific requirements. Our experienced team works closely with you at each step to ensure optimal results for your CAR-Treg development program. Below are the key services we provide, which can be customized based on your unique needs.

Biomarker Identification & ScFv Generation

Comprehensive target antigen identification and validation, followed by development of specific scFv.

Advanced screening of potential biomarkers for specific diseases.
Development and optimization of high-affinity scFv molecules.
Validation of target specificity and binding efficiency.
Customized antibody humanization services.

CAR Design & Construction

Expert design and optimization of CAR constructs tailored for Treg cell engineering.

Customized CAR vector design and optimization.
Selection of appropriate promoters and regulatory elements.
Integration of co-stimulatory domains.
Development of novel CAR architectures.

CAR Gene Packaging & Delivery

Efficient viral and non-viral delivery systems for optimal CAR gene transfer.

Lentiviral and retroviral vector production.
Optimization of transduction protocols.
Quality control and safety testing.
Alternative delivery method development.

CAR-Treg In Vitro Assay

Comprehensive in vitro testing to validate CAR-Treg functionality and efficacy.

Phenotype and function verification.
Cytokine production profiling.
Suppression activity assessment.
Stability and persistence evaluation.

Preclinical In Vivo Assay

Rigorous preclinical in vivo studies to evaluate safety and efficacy in animal models.

Development of appropriate disease models.
Biodistribution and trafficking studies.
Safety and toxicity assessment.
Long-term efficacy evaluation.

CAR-Treg Clinical Trial Support

Comprehensive support for clinical trial development and implementation.

Protocol development and optimization.
Regulatory compliance assistance.
GMP-compliant manufacturing support.
Clinical monitoring and data analysis.

Frequently Asked Questions

Can you help with the development of CARs targeting novel antigens specific to our disease of interest? What's your experience with non-standard targeting approaches?

Our extensive experience in CAR development encompasses both traditional and innovative targeting strategies. We provide comprehensive support for novel antigen targeting, beginning with sophisticated in silico analysis to identify promising candidates. Our platform includes the development and screening of custom scFv libraries, along with expertise in alternative targeting moieties such as single-domain antibodies, and artificial binding proteins. We optimize CAR constructs through iterative testing of different costimulatory domains and perform rigorous validation studies to ensure specificity and minimize off-target effects. Our team has successfully developed CARs for numerous novel targets across various disease indications, demonstrating our ability to tackle unique targeting challenges.

What are your requirements for starting materials when clients provide their own antibody samples for CAR-Treg development?

For client-provided antibody samples, we require a minimum concentration of 1 mg/mL in a stabilized buffer, preferably PBS or similar physiological buffer. The sample should be accompanied by initial characterization data including purity assessment, aggregation profile, and binding specificity information. We also request information about storage conditions and handling history. Before beginning the development process, we conduct our own quality assessment including sterility testing, endotoxin levels, and protein stability analysis to ensure suitability for CAR development. If needed, we can provide guidance on sample preparation and shipping conditions to maintain optimal antibody quality.

Can your platform accommodate complex CAR designs with multiple targeting domains or switchable systems?

Our platform has successfully developed and validated various complex CAR designs including dual-targeting CARs, switchable CAR systems, and logic-gated constructs. We utilize advanced molecular biology techniques and optimized cloning strategies to accommodate sophisticated designs while maintaining construct stability and expression efficiency. Our experience includes working with various combinations of co-stimulatory domains, incorporating safety switch systems, and developing regulatable expression systems. Each complex design undergoes thorough testing for structural integrity, expression stability, and functional validation to ensure all components work synergistically while maintaining Treg phenotype and function.

How do you handle the development of CAR-Tregs targeting tissue-specific antigens where the target may have limited expression or accessibility?

We employ a specialized approach for challenging tissue-specific targets, beginning with extensive in silico analysis of target expression patterns and accessibility. Our development process includes advanced screening methods to validate targeting even with limited antigen expression, utilizing sensitive detection systems and specialized binding assays. We develop customized potency assays that accurately reflect the intended tissue environment and employ tissue-specific migration assays to evaluate homing capacity. Our experience includes working with rare antigens and developing specialized protocols to ensure effective targeting while maintaining safety profiles.

Does your CAR-Treg platform support both autologous and allogeneic development approaches? What screening processes are in place for each?

Our platform is optimized for both autologous and allogeneic CAR-Treg development, with specific workflows designed for each approach. For autologous development, we implement comprehensive donor screening protocols and establish patient-specific cell characterization profiles. In allogeneic development, we utilize advanced HLA typing and screening methods to identify and select optimal donor populations, combined with sophisticated gene editing approaches to minimize alloreactivity. Our platform includes extensive characterization of TCR repertoire, evaluation of potential xenoreactive responses, and thorough assessment of expansion capacity to ensure consistent product quality regardless of the cellular source.

Reference

Zhang, Qunfang, et al. "Chimeric antigen receptor (CAR) Treg: a promising approach to inducing immunological tolerance." Frontiers in immunology 9 (2018): 2359. Distributed under Open Access license CC BY 4.0, without modification.