Active cxcr4+ immune cells and methods for their production and use

a technology of immune cells and active cxcr4+, applied in the field of immunotherapy and cancer, premalignant lesions, chronic infections, can solve the problems of out-running a robust immune response, significant obstacle to clinical effectiveness, and failure to detect tumors

Pending Publication Date: 2021-03-11
UNIV OF PITTSBURGH OF THE COMMONWEALTH SYST OF HIGHER EDUCATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The approach results in cells with increased CXCR4 surface expression and cytotoxicity, enabling them to effectively migrate to and target tumors, offering a potentially more efficient and cost-effective treatment for cancer and chronic infections.

Problems solved by technology

The development of tumors might be seen as a failure of immune surveillance.
However, not all tumors are naturally immunogenic, and even among those that are immunogenic, the uncontrolled rapid growth of a cancer may sometimes out-run a robust immune response.
Suboptimal homing of type-1 effector CD8+ T cells (typically referred to as cytotoxic T cells; CTLs) to tumors constitutes a significant obstacle to clinical effectiveness of the spontaneously-occurring and therapeutically-induced immunity against cancer.
However, modification of the CKR repertoire of CTLs to match chemokines expressed in tumor microenvironment (TEM) represents multiple obstacles.
Yet, genetic manipulation of cells for administration to a patient is difficult, time consuming, and expensive.
Similarly, activated natural killer (NK) cells are known to have type-1 homing pattern, This indicates that cx vivo or in vivo induction of T cells (or NK cells) with both the desirable cytotoxiclpro-inflammatory properties and the ability to enter typical tumor tissues will be difficult to achieve.

Method used

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  • Active cxcr4+ immune cells and methods for their production and use
  • Active cxcr4+ immune cells and methods for their production and use
  • Active cxcr4+ immune cells and methods for their production and use

Examples

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example 1

Materials and Methods

[0079]Taqman Analysis of mRNA Expression in Tumor

[0080]Four (4) mm punch biopsies of tumor tissues were placed in lysing Matrix E tubes Biologicals, Solon, Ohio) containing RUT buffer (RNeasy kit, Qiagen, Valencia, Calif.), and agitated using a FP120 homogenizer (MP Biologicals). Debris-free supernatant from the lysis matrix tubes were transferred into new tubes and total RNA was extracted using the RNeasy kit. One (1) μg of RNA extracted using this method was used for cDNA synthesis and 25-50 ng of subsequent cDNA was used to perform mRNA expression analysis by Taqman analysis on Step One Plus system (Applied Biosystems). All primers used for the analysis were standard primers purchased from Applied Biosystems.

Confocal Microscopy Analysis of Tumor Sections

[0081]Four (4) mm tumor punches, either untreated or treated, were embedded in OCT medium containing cryomolds and immediately frozen in 2-methyl-butane. Six (6) μm frozen sections of the tissues were made usi...

example 2

Uniform Expression of CXCL12 in Colorectal Tumors and Expression of High Levels of CXCR4 on Tumor-Infiltrating CD8+ T Cells

[0090]In order to identify the chemokine(s) uniformly expressed in cancer, RNA (ragman) analysis of tumors from 137 patients was performed. The results revealed a unique pattern of spontaneous CXCL12 expression, which was spontaneously expressed at high levels in colorectal cancer tissues from 137 patients, in sharp contrast to the effector T cell attracting chemokine CXCL10, which was either absent or expressed only at marginal levels (FIG. 1A). Confocal analysis revealed that both CD326 (EP-CAM) negative (non-.tumor) and positive (tumor) cells make CXCL12 (FIG. 1B). Moreover CD8+ tumor infiltrating lymphocytes (TILs) isolated from tumors, showed high expression of CXCR4, rather than CXCR3, indicating that CXCR4 can drive the attraction of immune cells, including CD8+ T cells (non-cytotoxic; data not shown), to cancer tissues (FIG. 1C).

example 3

[0091]Hypoxia and Small-Molecule HIF-1α Stabilizers Induce Double-pPsitive CXCR4+ / GZMB+CTL Cells in CTL / Th1-Driving Conditions

[0092]In accordance with the current paradigm of the regulation of chemokine receptors on pro-inflammatory and non / anti-inflammatory subsets of T cells [Sallusto, F., et al. 1998. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J. Exp. Med. 187:875-883; Sallusto, F., and Mackay, C. R. 2004. Chemoattractants and their receptors in homeostasis and inflammation. Curr. Opin. Immunol. 16:724-731; Sallusto, F., Mackay, C. R., and Lanzavecchia, A. 1997. Selective expression of the eotaxin receptor CCR3 by human T helper 2 cells. Science 277:2005-2007; Sallusto, F., Mackay, C. R., and Lanzavecchia, A. 2000. The role of chemokine receptors in primary, effector, and memory immune responses. Annu. Rev. Immunol. 18:593-620; Galli, G., et al. 1998. Enhanced HIV expression during Th2-oriented responses explained by the op...

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Abstract

Provided herein are active CXCR4+ CD8; T cells, active CXCR4+ type-1 CD4+ T cells and active CXCR4+ NK cells and populations of those cells, methods for making active CXCR4+ T cells and NK cells and populations of those cells, and methods for using active CXCR4+ T cells and NK cells and populations of those cells for the treatment of cancer, precartcerous conditions and chronic infections.

Description

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0001]This invention was made with government support under CA132714 and CA121973 awarded by the National Institutes of Health. The government has certain rights in this invention.BACKGROUND OF THE INVENTION1) Field of the Invention[0002]The fields of the invention are immunology, immunotherapy and treatment of cancer, premalignant lesions and chronic infections,2) Description of Related Art[0003]Cancer (or neoplastic disease) is the name we give to a mass of cells that grows in an abnormal, unregulated way and that ultimately overwhelms a body system or organ. A “tumor” refers to any abnormal growth of cells and can be harmless or dangerous. A harmless tumor is called benign and does not contain cancerous cells whereas a dangerous tumor is called malignant because it contains cancerous cells.[0004]Cancers are further divided into carcinomas, sarcomas, leukemias or lymphomas, according to the cell types that are involved...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): C12N5/0783A61K35/17C12N5/071
CPCC12N5/0638A61K35/17C12N5/0602C12N2501/60C12N5/0637C12N2500/02C12N2501/2312C12N5/0636A61K39/464839A61K39/46433A61K39/4621A61K39/46446A61K39/4611A61K39/464421
InventorKALINSKI, PAWELMUTHUSWAMY, RAVIKUMAR
OwnerUNIV OF PITTSBURGH OF THE COMMONWEALTH SYST OF HIGHER EDUCATION