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Compositions, methods and products comprising human papillomavirus for detecting and treating a cancer

a technology of human papillomavirus and cancer, applied in the field of products, compositions, methods and equipment for identifying cancers and precancerous cellular changes, can solve the problems of breast cancer not being detected in all cases, breast cancer cannot be currently prevented, and the risk is increasing

Inactive Publication Date: 2006-02-09
HERMONAT PAUL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention provides methods, compositions, and products for screening and treating cancers and pre-cancerous cellular changes, particularly breast cancer and pre-cancerous breast changes. These methods involve amplifying a sample from a biopsy taken from a patient and probing it for the presence of an HPV sequence. The invention also provides compositions and methods for inhibiting the expression of HPV genes or proteins. The technical effects of the invention include improved screening and treatment for cancer and pre-cancerous cellular changes, particularly breast cancer and pre-cancerous breast changes."

Problems solved by technology

All women are at risk for breast cancer, with this risk increasing as a woman ages.
Breast cancer cannot currently be prevented.
However, not all breast cancers are currently detected at this early stage.
It appears that whatever tissue site HPVs are known to infect, they cause pathology.
However, there is a significant risk that this higher than normal active cell growth may become an outright malignancy.
However, as recent studies have indicated, the relationship between HPV infection and breast cancer is controversial.
However, of the control study of eight patients having breast cancer diagnosed before the CIN III lesions, none had HPV positive breast carcinomas.

Method used

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  • Compositions, methods and products comprising human papillomavirus for detecting and treating a cancer
  • Compositions, methods and products comprising human papillomavirus for detecting and treating a cancer
  • Compositions, methods and products comprising human papillomavirus for detecting and treating a cancer

Examples

Experimental program
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example 1

Analysis of Breast Cancer Tissue for Presence of HPV Via PCR

[0087] In the present example, total DNA was isolated from breast cancer tissues and analyzed for the presence of HPV by use of PCR amplification. The amplification targeted the L1 gene and was broad spectrum, thus allowing for amplification of many different HPV types.

[0088] Patients, Breast Cancer Specimens, and DNA Isolation

[0089] 17 women with breast cancer receiving examinations and treatment at the University of Arkansas for Medical Sciences (UAMS) from May 1999 to October 1999. Portions of needle biopsy tissue was fixed in a Phosphate Buffered Saline: Ethanol (1:1) solution soon after they were acquired. All of the specimens were stored at −80° C. The specimens were processed by grinding, and total cellular DNA was isolated from the specimens by pelleting and resuspending them in lysis buffer (0.5 mg / ml Proteinase K, 0.5% SDS, 0.5 mM EDTA, 0.5 mM Tris-HCL, pH7.4). After incubation overnight at 37° C., the total ce...

example 2

Construction of the AAV / E6 / Neo Genome, Generation of Virus Stocks, and Titering of Virus Stocks

[0103] The AAV / E6 / Neo genome was constructed as a plasmid, in a similar manner to the construction of the AAV / GM-CSF / Neo viral genome as described by Liu (Liu. Y., et al., J. Inf. Cytok. Res. 2000; 20:21-30), incoporated herein by reference. However, instead of the GM-CSF gene, the HPV-16 E6 open reading frame was cloned by PCR amplification using Pfu polymerase and ligated into the vector. A structural map of the AAV / E6 / Neo vector used in this study is shown in FIG. 8A. In this construct the E6 gene is expressed from the AAV p5 promoter, which is known to be active in DC. An AAV / E7 / Neo vector was also made in this study (not shown). In the E7 construct, the E7 gene is expressed from the AAV p5 promoter.

[0104] High titer rough (non-purified) rAAV virus stocks were generated in a two-step process, using the complementor plasmid ins96-0.8, and titered as described previously by Hermonat et...

example 3

Higher CD8 / CD4 and Lower CD56 / CD8 Cell Ratios Result with AAV-Mediated Pulsing / Priming

[0130] The makeup of the T cell populations, which resulted from AAV-transduction or protein lipofection, was observed. An effective CTL response, while requiring CD8+ T cells as an effector of lysis, also requires CD4+ helper T cells. Flow cytometric analysis was used to determine the phenotype of the population of the lysate, GST-E6 pulsed, and AAV / E6 / Neo pulsed T cell populations.

[0131]FIG. 14A shows the CD8 and CD4 prevalence within the primed population resulting from three different techniques as indicated (on the right), as well as an FL1-H, FL2-H control (left). FIG. 14B shows the CD56 and CD8 ratios in the same experimental situations as A.

[0132] As shown in FIG. 14A, in the mock case, one sees a normal ratio of CD8 to CD4 postive cells (1.21:1). In the GST-E6 pulsing case, the D8 / CD4 ratio remains the same (1.23:1). In sharp contrast in the AAV / E6 / Neo pulsed case, the ratio of CD8 / CD4 ...

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Abstract

Methods for screening a patient for a cancer wherein the methods comprise detecting an HPV in a biopsy from a patient are disclosed. Also disclosed are compositions and products for screening and for treating cancer in a patient, as well as methods of treating a patient afflicted with a cancer.

Description

BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention relates to products, compositions, methods and apparatus for identification of cancers and pre-cancerous cellular changes. In another aspect, the present invention relates to products, compositions, methods and apparatus for identification of breast cancers or pre-cancerous cellular changes in breast tissues. In even another aspect, the present invention relates to products, compositions, methods and apparatus for treatment of cancers and pre-cancerous cellular changes. In still another aspect, the present invention relates to products, compositions, methods and apparatus for treatment of breast cancer and pre-cancerous cellular changes in breast tissues. [0003] 2. Description of the Related Art [0004] Breast cancer is the most common form of cancer in women in the United States. It is estimated that in the year 2000, 182,800 new cases of female invasive breast cancer will be diagnosed, and 40,...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C12P19/34A61B10/00
CPCG01N2333/025A61B10/0041
Inventor HERMONAT, PAULKLIMBERG, V.LIU, YONG
Owner HERMONAT PAUL
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