Site-Specific In Vivo T Cell Engineering for Targeted Integration

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Solution Overview

Problem

Current cancer immunotherapy relies on cumbersome and expensive ex vivo manipulations of T cells, which are time-consuming and require specialized facilities, limiting its accessibility and affordability for most patients.

Innovation Solution

A system for site-specific in vivo engineering of T cells using a site-specific nuclease to target the integration of exogenous nucleic acid sequences into desired genomic loci, such as the TRAC and TRBC loci, via homologous recombination, using AAV vectors and site-specific nucleases like ARCUS, enabling precise and scalable T cell engineering.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If ex vivo T cell engineering is used to achieve precise T cell modification, then manufacturing precision is improved, but device complexity and treatment cost increase

Engineering Contradiction:

ImproveT cell modification precisionVSAvoidspecialized facilities and expertise requirements

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent enables T cells to perform self-engineering in vivo by expressing site-specific nucleases and homologous recombination machinery within the patient's body, eliminating the need for complex ex vivo manipulation facilities and specialized expertise while maintaining precise targeted integration at the TRAC locus

Inventive Principle:

Principle #25Self-service

2Manufacturing precision

If ex vivo T cell engineering is used to achieve precise T cell modification, then manufacturing precision is improved, but loss of time increases

Engineering Contradiction:

ImproveT cell modification precisionVSAvoidtreatment timeline

Core Design Contradiction:

Manufacturing precisionVSLoss of time

Solution Approach 1:

The patent inverts the conventional ex vivo engineering approach by performing T cell modification in vivo within the patient's body. This reversal eliminates time-consuming steps including cell collection, transport to specialized centers, viral vector manufacturing, and cell reinfusion logistics, while maintaining precise targeted integration through site-specific nuclease expression and homologous recombination

Inventive Principle:

Principle #13The other way round (Inversion)

3Ease of manufacture

If commonly used integration techniques are used to insert CAR genes, then ease of manufacture is improved, but reliability worsens due to random integration and variegated expression

Engineering Contradiction:

Improveintegration process simplicityVSAvoidintegration precision and expression consistency

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

The patent applies local quality by using site-specific nucleases to create targeted double-strand breaks at the TRAC locus, followed by homologous recombination with donor templates containing homology arms. This localized precise integration mechanism ensures consistent CAR expression and eliminates random integration events, while the modular nuclease-donor vector system maintains manufacturing simplicity

Inventive Principle:

Principle #3Local quality

4Productivity

If site-specific in vivo engineering is used to reduce treatment cost and timeline, then productivity is improved, but manufacturing precision may worsen without proper targeting

Engineering Contradiction:

Improvetreatment scalability and affordabilityVSAvoidnucleic acid integration precision

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent uses site-specific nucleases as intermediaries to mediate precise in vivo T cell engineering. These nucleases recognize specific DNA sequences at the TRAC locus, create targeted double-strand breaks, and facilitate homologous recombination with donor templates. This intermediary mechanism ensures high integration precision while enabling scalable in vivo treatment that reduces costs and timelines compared to ex vivo approaches

Inventive Principle:

Principle #24Intermediary (Mediator)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach reduces treatment costs and timelines, eliminates the need for pre-conditioning chemotherapy, and prevents graft-versus-host disease, providing a scalable and affordable immunotherapy for a wide range of patients.

Implementation Method 1

targeting the integration of exogenous nucleic acid sequences into a desired target locus... integration to the target locus by homologous recombination

Methodology Applied

Scientific EffectHomologous recombination:

Data Source

PatentUS20250222138A1Site-specific in vivo t cell engineering, systems, compositions and methods thereof

Publication Date: 2025.07.10 RAMOT AT TEL AVIV UNIVERSITY LTD

AI summary

The present disclosure relates to immunotherapy. In more specific embodiments, the present disclosure provides systems, compositions, methods and uses of viral vectors comprising nucleic acid sequence of interest that encodes at least one therapeutic product, and a nucleic acid sequence encoding at least one nuclease, for in vivo targeted insertion of the nucleic acid sequence of interest into a target locus within at least one cell of the T lineage.