Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Cll1-specific multi-chain chimeric antigen receptor

Inactive Publication Date: 2018-01-04
CELLECTIS SA
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a novel approach for immunotherapy of CLL1 positive cancer cells, through the use of multi-chain CARs (mcCARs) that specifically target and kill these cells. The mcCARs are designed to have less toxic side effects, including cytokine release, and can be used in combination with classical anti-cancer drugs. The resulting engineered T-cells display reactivity against CLL1 positive cells and are useful for immunotherapy. The invention also provides allogeneic immune cells that can be used as off-the-shelf therapeutic products.

Problems solved by technology

However, this approach has so far proven efficiency only with respect to patients with acute lymphoblastic leukemia (ALL) by targeting malignant B cells bearing the antigen CD19 (Porter, D. L. et al.
Induction treatments for acute myeloid leukemia (AML) have remained largely unchanged for nearly 50 years and AML remains a disease of poor prognosis.
Meanwhile, induction treatments for acute myeloid leukemia (AML) have remained largely unchanged for nearly 50 years and AML remains a disease of poor prognosis.
However, none of these antibodies have been reported to date as being tested in clinical trials as therapeutic antibodies.
Monoclonal antibodies have often been used to treat lymphomas, but their use in leukemias has been more limited.
However, no therapeutic effects could be attributed to the CCL-1 targeting peptide per se.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Cll1-specific multi-chain chimeric antigen receptor
  • Cll1-specific multi-chain chimeric antigen receptor
  • Cll1-specific multi-chain chimeric antigen receptor

Examples

Experimental program
Comparison scheme
Effect test

example 1

Multi-Chain CARs

[0674]Multi-chain CARs targeting the CLL1 antigen were designed based on the high affinity receptor for IgE (FcεRI) such as depicted in FIG. 2 to FIG. 4. The FcεRI expressed on mast cells and basophiles triggers allergic reactions. It is a tetrameric complex composed of a single a subunit, a single β subunit and two disulfide-linked γ subunits. The a subunit contains the IgE-binding domain. The β and γ subunits contain ITAMs that mediate signal transduction. In every multi-chain CAR, the extracellular domain of the FcRα chain was deleted and replaced by the respective scFv referred to μ ln Table 5 respectively and the CD8α hinge (SEQ ID NO: 2) and the ITAM of the FcRβ chain and / or the FcRγ chain was deleted. The resulting constructions had the structure detailed in table 6.

example 2

n of Anti-CLL1 mcCARs in Human T Cells

[0675]Primary T-Cell Cultures

[0676]T cells were purified from Buffy coat samples provided by EFS (Etablissement Francais du Sang, Paris, France) using Ficoll gradient density medium (Ficoll Paque PLUS / GE Healthcare Life Sciences). The PBMC layer was recovered and T cells were purified using a commercially available T-cell enrichment kit (Stem Cell Technologies). Purified T cells were activated in X-Vivo™-15 medium (Lonza) supplemented with 20 ng / mL Human IL-2 (Miltenyi Biotech), 5% Human Serum (Sera Laboratories), and Dynabeads Human T activator CD3 / CD28 at a bead:cell ratio 1:1 (Life Technologies). After activation cells were grown and maintained in X-Vivo™-15 medium (Lonza) supplemented with 20 ng / mL Human IL-2 (Miltenyi Biotec) and 5% Human Serum (Sera Laboratories).

[0677]Models of AML and Clorofarabine, Fludarabine or Cytarabine Resistant AML

[0678]Originally, an AML-positive cell line, such as MOLM13 cell line, has been established from the ...

example 3

tion of T Cells Transiently Expressing the Anti-CLL1 mcCARs Following Coculture with Target Cells

[0688]24 hours post electroporation, human T cells engineered using polycistronic mRNAs encoding the multi-chain CARs were co-cultured with target (Daudi) or AML cell line control cells for 6 hours. The CD8+ T cells were then analyzed by flow cytometry to detect the expression of the degranulation marker CD107a at their surface. This experiment aims to check that the human CD8+ T cells expressing the CLL1 multi-chain CARs degranulate in coculture with CLL1 expressing target cells but not in coculture with control cells.

[0689]Degranulation Assay (CD107a Mobilization)

[0690]T-cells were incubated in 96-well plates (40,000 cells / well), together with an equal amount of cells expressing or not the CLL1 protein. Co-cultures were maintained in a final volume of 100 μl of X-Vivo™-15 medium (Lonza) for 6 hours at 37° C. with 5% CO2. CD107a staining was done during cell stimulation, by the addition...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Fractionaaaaaaaaaa
Fractionaaaaaaaaaa
Lengthaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a new generation of chimeric antigen receptors (CAR) referred to as multi-chain CARs, which are made specific to the antigen CLL1. Such CARs aim to redirect immune cell specificity and reactivity toward malignant cells expressing the tumor antigen CLL1. The alpha, beta and gamma polypeptides composing these CARs are designed to assemble in juxtamembrane position, which forms flexible architecture closer to natural receptors, that confers optimal signal transduction. The invention encompasses the polynucleotides, vectors encoding said multi-chain CAR and the isolated cells expressing them at their surface, in particularly for their use in immunotherapy. The invention opens the way to efficient adoptive immunotherapy strategies for treating cancer, especially leukemia.

Description

FIELD OF THE INVENTION[0001]The present invention relates to a new generation of chimeric antigen receptors (CAR) referred to as multi-chain CARs, which are made specific to the antigen CLL1. Such CARs aim to redirect immune cell specificity and reactivity toward malignant cells expressing the tumor antigen CLL1. The polypeptides composing these CARs are designed to assemble in juxtamembrane position, which forms flexible architecture closer to natural receptors, that confers tunable signal transduction. The invention encompasses the polynucleotides, vectors encoding said multi-chain CAR and the isolated cells resulting from their heterologous expression in immune cells, in particularly for their use in immunotherapy. The invention opens the way to efficient adoptive immunotherapy strategies for treating cancer, especially acute myeloid leukemia (AML).BACKGROUND OF THE INVENTION[0002]Adoptive immunotherapy, which involves the transfer of antigen-specific T cells generated ex vivo, i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/28C12N5/00C07K16/30A61K39/00A61K35/17C12N5/0783C07K14/725
CPCC07K16/2866A61K35/17A61K39/0011C07K14/7051C07K2317/622C12N5/0093C12N5/0638A61K2039/5158A61K2039/64C07K16/3061C07K16/2803C07K2317/24C07K2319/03C12N2510/00C07K14/70596C12N5/0636A61P35/00C07K2319/02C07K2319/33A61K39/4611A61K39/4631A61K39/4644C07K2319/00A61K2039/6056A61P35/02A61P43/00C07K16/28C07K14/70517C07K14/70535C07K14/70578C07K16/2887A61K39/39558G01N15/1031G01N15/14C07K2317/53C07K2317/56A61K2039/505A61K2039/5156G01N2015/016G01N2015/1006G01N2015/1028G01N15/149A61K39/395C07K19/00C12N5/10C12N15/63A61K39/001176
Inventor SMITH, JULIANNEVALTON, JULIENDUCHATEAU, PHILIPPEJUILLERAT, ALEXANDRERAJPAL, ARVINDSASU, BARBRA JOHNSON
Owner CELLECTIS SA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com