In vivo CAR-T therapy represents a transformative paradigm in adoptive cell therapy—eliminating ex vivo manufacturing through direct in situ generation of functional CAR T cells within the patient.
Alfa Cytology offers an integrated preclinical CRO solution for in vivo CAR-T development—bridging molecular design, vector optimization, and animal model validation to accelerate translational proof-of-concept for next-generation cellular immunotherapies.
Introduction to In Vivo CAR-T
The in vivo CAR T therapy simplifies ex vivo CAR T method by systemic administration of the CAR gene editing construct enveloped in viral vectors or nanoparticles. These carriers specifically target T cells to unload gene editing cargo, thus inducing the expression of the CAR construct on the T cell surface. The resulting CAR T cells can then specifically detect cancer cells, thus activating themselves and expanding to effectively eliminate cancer cells in the bloodstream or malignant tumours.
Fig.1 Process of ex vivo CAR T therapies.
Fig.2 Process of the in vivo CAR T therapies.
Platform Core — In Vivo CAR-T Engineering
The in vivo CAR-T platform redefines adoptive cell therapy by generating functional CAR-T cells directly within the body, removing the need for ex vivo manufacturing. Current development converges on two complementary delivery strategies, each designed to enable precise, safe, and efficient T-cell reprogramming in vivo.
Targeted Viral Vector Systems
Viral vectors remain the most mature approach for in vivo CAR-T generation owing to their high transduction efficiency and durable gene integration.
They can be surface-engineered to achieve T-cell specificity through display of scFvs (e.g., anti-CD3, anti-CD4, anti-CD8) or bispecific ligands on the viral envelope. Key types include:
Targeted Nanoparticle-Based Systems
Non-viral nanocarriers offer a flexible, transient, and scalable alternative for in vivo CAR-T generation. By encapsulating mRNA or plasmid DNA encoding CAR constructs and decorating the surface with T-cell-specific antibodies or ligands, these systems enable safe, non-integrating, and repeatable T-cell transfection. Major types include:
Comparative Features: Targeted Viral Vectors vs. Targeted Nanoparticle Vectors
| Feature | Targeted Viral Vectors | Targeted Nanoparticle Vectors |
| Representative Types | Lentivirus, Retrovirus, AAV | LNPs, Polymeric Nanocarriers, Exosomes |
| Delivered Cargo | DNA (integrating or episomal) | mRNA, plasmid DNA, or RNP (non-integrating) |
| T-Cell Targeting Strategy | Envelope modification with anti-CD3/CD4/CD8 scFvs or bispecific ligands | Surface conjugation with anti-CD3/CD5/CD8 antibodies or peptides |
| Gene Expression Profile | Long-term, stable integration | Transient (days to weeks), tunable by mRNA half-life |
| Delivery Efficiency | Very high (>60–80% in preclinical models) | Moderate (5–20%, formulation-dependent) |
| Immunogenicity | Viral components may trigger innate/adaptive immune responses | Low; chemically defined lipids/polymers reduce immune activation |
| Safety | Risk of insertional mutagenesis and inflammatory reactions; replication-competent virus must be controlled | No insertional risk; some cationic materials may induce toxicity |
| Production Complexity | Cell-based, multi-step manufacturing; costly QC | Scalable microfluidic or chemical synthesis; low cost |
| Stability & Storage | Requires cold-chain logistics and sterile handling | Stable; can be lyophilized and stored at room or refrigerated conditions |
| Ideal Application | Durable hematologic CAR-T generation | Rapid, reversible, and safe CAR-T expression for solid tumors |
In Vivo CAR-T Development Workflow
Applications of In Vivo CAR-T
Oncology Applications
Infectious and Autoimmune Diseases
Regenerative and Fibrotic Disorders
Why Choose Us?
FAQs
It eliminates patient-specific cell collection and culture, reducing production time from weeks to days and lowering costs while maintaining therapeutic potency.
No. They are complementary modalities — viral vectors for durable integration and LNP for transient expression or solid-tumor testing. Both can be evaluated within Alfa Cytology's platform.
Third-generation self-inactivating vectors are used; replication-competent virus testing and integration-site mapping are included in QC to avoid insertional mutagenesis.
Yes. LNP systems are non-integrating and immunologically tolerant, allowing repeat administration for boosted response or multi-antigen targeting.
CD34+ and PBMC humanized mice for hematologic studies, and orthotopic or systemic tumor models for solid tumor testing with flow, qPCR, and bioluminescence tracking.
Alfa Cytology, a pioneer in alfaCAR-TTM therapy, provides integrated preclinical solutions to accelerate next-generation cancer immunotherapy development. Our technology platform combines advanced alfaCAR-TTM design, precision engineering, and multi-dimensional characterization to overcome key challenges and expedite therapy translation.
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