Engineered cells and uses thereof

A cell, immune cell technology, applied in the field of engineered cells and their uses, can solve problems such as off-target toxicity, release syndrome, and unwanted cytokines

Pending Publication Date: 2021-11-30
NANJING LEGEND BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these therapies suffer from a number of drawbacks, including off-target toxicity and unwanted cytokine release syndrome in treated subjects
This and other side effects can further exacerbate the inflammatory response, organ failure, and in extreme cases, death

Method used

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  • Engineered cells and uses thereof
  • Engineered cells and uses thereof
  • Engineered cells and uses thereof

Examples

Experimental program
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Effect test

example

[0193] Various aspects of the present disclosure are further illustrated by the following non-limiting examples.

example 1

[0194] Example 1: Design of a TCR+ CAR construct containing only a co-stimulatory domain

[0195] This example describes the design of an exemplary TCR+ CAR construct comprising only a co-stimulatory domain. Twelve constructs were designed, each comprising a polynucleotide as follows:

[0196] Construct 1 (anti-CLL-1 TCRε+anti-CD33 co-stimulatory CAR): polynucleotide contains CD3ε leader peptide from N-terminus to C-terminus-anti-CLL-1 sdAb-(G4S) 3 -CD3ε-T2A-CD8 leader peptide-anti-CD33 sdAb-CD8 hinge-CD8 transmembrane-CD27 co-stimulatory domain;

[0197] Construct 2 (anti-CLL-1 TCRδ+anti-CD33 co-stimulatory CAR): polynucleotide contains CD3δ leader peptide from N-terminus to C-terminus-anti-CLL-1 sdAb-(G4S) 3 -CD3δ-T2A-CD8 leader peptide-anti-CD33 sdAb-CD8 hinge-CD8 transmembrane-CD27 co-stimulatory domain;

[0198] Construct 3 (anti-CLL-1 TCRγ + anti-CD33 co-stimulatory CAR): polynucleotide contains CD3γ leader peptide from N-terminus to C-terminus-anti-CLL-1 sdAb-(G4S) ...

example 2

[0222] Example 2: Viral transfection and virion production

[0223]To generate viral particles comprising polynucleic acids encoding any of the systems disclosed herein, a mixture of lentiviral packaging plasmids including pMDLg / pRRE (Addgene #12251), pRSV-Rev (Addgene #12253) and pMD2.G (Addgene #12259) was mixed with PLVX-EF1A (including target system) vector was premixed with polyetherimide (PEI) at a pre-optimized ratio, mixed well and incubated at room temperature for 5 minutes. Add the transfection mixture dropwise to 293-T cells and mix gently. Incubate the transfected 293-T cells at 37°C and 5% CO 2 Incubate overnight. Twenty-four hours after transfection, the supernatant was collected and centrifuged at 500 g for 10 min at 4°C to remove any cell debris. The centrifuged supernatant was filtered through a 0.45 μm PES filter, and the virus supernatant was concentrated after ultracentrifugation. After centrifugation, carefully discard the supernatant and wash the virus...

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PUM

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Abstract

A system for inducing activity of immune cells comprising a chimeric antigen receptor, a T cell receptor, and various combinations thereof, an isolated host cell expressing the system, an antigen-specific immune cell comprising the system and methods of immunotherapy are provided.

Description

Background technique [0001] Effector cell activity generally involves the binding of a ligand to a receptor bound to the effector cell membrane. This interaction between the ligand and the membrane-bound receptor (usually via its extracellular binding domain) can lead to conformational and / or chemical modification of the receptor itself, which in turn can generate a cascade of events that accumulate leading to effector cell activation. Intracellular signaling. Attempts to exploit this interaction to develop immune cell therapies against cancer cells have shown promising efficacy against hematological malignancies. However, these therapies suffer from a number of drawbacks, including off-target toxicity and unwanted cytokine release syndrome in treated subjects. This and other side effects can further exacerbate the inflammatory response, organ failure and, in extreme cases, death. Contents of the invention [0002] In view of the foregoing, there is a great need for alter...

Claims

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Application Information

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IPC IPC(8): C12N5/10C07K19/00A61K35/17A61P35/00A61P31/00A61P31/12
CPCA61P31/12A61P35/00A61P31/00C07K2319/03C07K14/7051A61K2039/572C12N5/0636C12N2510/00C12N2501/515A61K39/4611A61K39/4632A61K39/464402A61K39/4631A61K2239/48A61K39/464411A61K35/17C07K16/28C07K16/2866C07K16/30C07K2319/00C12N15/86C12N2501/2302C12N2740/15043
Inventor 涂中原张贺忠张亚峰武术
Owner NANJING LEGEND BIOTECH CO LTD
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