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

DNA composition against tumor stromal antigen FAP and methods of use thereof

A composition and tumor technology, applied in biochemical equipment and methods, DNA/RNA vaccination, complete cell/virus/DNA/RNA components, etc., can solve the problem of high production cost and achieve the effect of stable expression

Inactive Publication Date: 2009-08-12
THE SCRIPPS RES INST
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Subunit vaccines have the advantages of being stable, safe, and chemically well-defined; however, they are too expensive to produce

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
  • DNA composition against tumor stromal antigen FAP and methods of use thereof
  • DNA composition against tumor stromal antigen FAP and methods of use thereof
  • DNA composition against tumor stromal antigen FAP and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Example 1. Preparation of DNA composition 1 encoding murine FAP.

[0120] will encode murine FAP (SEQ ID NO: 4, Figure 10 ) cDNA (SEQ ID NO: 3, Figure 9 ; provided by Dr.J.D.Cheng) subcloned into the EcoRI restriction site of pcDNA3.1 / V5-His-TOPO vector (Invitrogen, San Diego, California) to obtain the eukaryotic expression vector pcDNA3.1-FAP (pFAP) (see figure 1 , sub-figure A). Proper expression of the 95 kDa FAP protein from the above vector was demonstrated by Western blotting of lysates from transiently transfected CT26 colon cancer cells, which do not express FAP themselves ( figure 1 , sub-figure B). Transiently transfected D2F2 breast cancer cells were also treated in the same way. For transient transfections, CT26 and D2F2 cells were electroporated as previously described (see Loeffer et al., FASEB, J. 2001, 15:758-767, incorporated herein by reference). FAP protein expression was demonstrated by Western blotting using a polyclonal rabbit anti-mouse FA...

Embodiment 2

[0123] Example 2. Preparation of DNA composition 2 encoding murine FAP, murine IL-2, and murine CCL21.

[0124] By the same general method described in Example 1, cDNA (DNA SEQ ID NO: 5) encoding murine IL-2 from ATCC, accession number 39892, Manassas, Virginia, and from Invitrogen, San Diego, California The cDNA of murine CCL21a (DNA SEQ ID NO: 7) was subcloned into the murine pFAP vector of Example 1. RE88 Salmonella typhimurium was transformed with the generated vector (pFAP / IL-2 / CCL21 ) by the method described in Example 1 to provide a DNA composition 2 solution comprising the following DNA construct: The DNA constructs operably encode murine FAP, murine IL-2, and murine CCL21a, and are incorporated into an attenuated Salmonella typhimurium vector.

Embodiment 3

[0125] Example 3. Evaluation of DNA compositions of the invention in murine models of colon and breast cancer.

[0126] Oral inoculation, tumor cell challenge and treatment with doxorubicin. For experiments in the prophylactic setting, the 9 Salmonella typhimurium (S.typhimurium) (AroA - dam - BALB / c mice (n=8) were inoculated 3 times by oral gavage of about 100 μl PBS at about 1 week intervals, the Salmonella typhimurium (S.typhimurium) was transformed with a vector plasmid encoding murine FAP Salmonella typhimurium (DNA composition 1 of Example 1), Salmonella typhimurium (DNA composition 2 of Example 2) transformed with vector plasmids encoding murine FAP, IL-2 and CCL21, or the DNA composition 2 of Example 1 For control vaccines, the above process was performed by using the method described in Niethammer, et al., Nat. Med., 2002, 8: 1369-1375. About 10 days later, by subcutaneous (s.c.) injection of about 3×10 4 CT26 colon cancer cells or by orthotopic (o.t.) injection...

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

No PUM Login to View More

Abstract

A DNA composition effective for eliciting an immune response against tumor cells and tumor metastases comprising a DNA construct that encodes for at least one epitope of fibroblast activation protein (FAP), which is expressible in immune cells, and which is incorporated in a pharmaceutically acceptable carrier. The composition can encode a single epitope of FAP, a polypeptide comprising two or more epitopes of FAP, the entire FAP protein, or any portion thereof that will elicit the desired immune response. In one preferred embodiment, the composition also includes a DNA construct that encodes an immune effector protein, such as a cytokine. The DNA composition of the invention can be used alone or in combination with a chemotherapeutic agent to treat diseases such as tumors and tumor metastases.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Patent Application Serial No. 60 / 815,316, filed June 21, 2006, which is incorporated herein by reference. [0003] government rights [0004] This invention was made with Government support under Grant No. CA83856 awarded by the National Institutes of Health and Grant No. BC031079 awarded by the Department of Defense. The government has certain rights in this invention. field of invention [0005] The present invention relates to deoxyribonucleic acid (DNA) compositions encoding suitable molecules effective to stimulate an immune response against stromal fibroblasts in tumors. In more detail, the present invention relates to DNA compositions encoding a matrix antigen known as fibroblast activation protein (FAP). The present invention also relates to methods of using the DNA composition to inhibit tumor growth and tumor metastasis, and to increase cellular uptake of...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K39/00
CPCA61K39/0005A61K2039/53A61K39/0011A61K2039/585A61P35/00A61P35/04A61P37/04A61P43/00
Inventor R·A·雷斯菲尔德M·勒夫勒J·A·克吕格尔A·G·尼萨默
Owner THE SCRIPPS RES INST
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