Analysis of genomic DNA, RNA, and proteins in exosomes for diagnosis and theranosis

Patent metadata

Field	Value
Publication number	US-9921223-B2
Application number	US-201415101857-A
Country	US
Kind code	B2
Filing date	Dec 4, 2014
Priority date	Dec 4, 2013
Publication date	Mar 20, 2018
Grant date	Mar 20, 2018

Abstract

The present invention provides that exosomes from human body fluid samples contain double stranded genomic DNA that spans all chromosomes and may be used to determine the mutation status of genes of interest in diseases, such as cancer. Furthermore, the present invention provides the use of exosomes to produce therapeutic proteins and for their use in therapy as well as the detection of cancer cell-derived exosomes to diagnose cancer and monitor therapeutic response.

First claim

The invention claimed is:

1. A method of isolating cancer cell-derived exosomes comprising:
(a) obtaining a body fluid sample from a cancer patient;
(b) isolating an exosomes fraction of the body fluid sample; and
(c) isolating exosomes from the exosomes fraction based on surface expression of glypican 1, thereby isolating cancer cell-derived exosomes.

2. The method of claim 1 , further comprising isolating genomic double-stranded DNA, RNA, or proteins from the cancer cell-derived exosomes.

3. The method of claim 2 , further comprising detecting a DNA, RNA, or protein isolated from the cancer cell-derived exosomes.

4. The method of claim 1 , wherein the isolating of step (b) or (c) comprises immunomagnetic capture, adhesion-based sorting, magnetic-activated sorting, or fluorescence-activated sorting (FACS).

5. The method of claim 1 , further comprising quantifying the number of cancer cell-derived exosomes in the patient.

6. The method of claim 1 , further comprising genotyping the cancer cell-derived exosomes.

7. The method of claim 1 , wherein the body fluid sample is lymph, saliva, urine, cerebrospinal fluid, bone marrow aspirates, eye exudate/tears, or serum.

8. The method of claim 1 , wherein the cancer is a breast cancer, lung cancer, head & neck cancer, prostate cancer, esophageal cancer, tracheal cancer, brain cancer, liver cancer, bladder cancer, stomach cancer, pancreatic cancer, ovarian cancer, uterine cancer, cervical cancer, testicular cancer, colon cancer, rectal cancer or skin cancer.

9. The method of claim 1 , wherein isolating exosomes from the exosomes fraction based on surface expression of glypican 1, comprises isolating the exosomes using a glypican 1-binding antibody.

10. The method of claim 3 , comprising isolating dsDNA from the cancer cell derived exosomes.

11. The method of claim 3 , comprising sequencing DNA from the cancer cell derived exosomes.

12. The method of claim 10 , comprising determining a mutation in at least one gene encoded by the dsDNA.

Assignees

• Univ Texas

Inventors

• Kalluri Raghu
• Melo Sónia

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• G01N33/57585 Primary

  involving compounds identifiable in body fluids · CPC title

• C12Q2600/158

  Expression markers · CPC title

• A61K9/5068

  Cell membranes or bacterial membranes enclosing drugs (liposomes with additional exogenous lipids A61K9/127) · CPC title