PEG-Prom mediated surface expression of avidin/streptavidin

Patent metadata

Field	Value
Publication number	US-9994865-B2
Application number	US-201414895125-A
Country	US
Kind code	B2
Filing date	Jun 4, 2014
Priority date	Jun 4, 2013
Publication date	Jun 12, 2018
Grant date	Jun 12, 2018

Abstract

The presently disclosed subject matter relates to compositions and methods directed to cancer theranostic nucleic acid constructs that permit simultaneous cancer-specific viral replication, expression of a diagnostic gene product, and expression of a therapeutic gene.

First claim

That which is claimed:

1. A nucleic acid construct comprising a first promoter operably linked to a gene encoding avidin or streptavidin comprising a membrane localization signal and a transmembrane domain, and a second promoter operably linked to a gene encoding a therapeutic agent, wherein the first promoter is cancer-selective, and wherein the construct comprises a sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18.

2. The nucleic acid construct of claim 1 , wherein the second promoter is a cancer-selective promoter.

3. The nucleic acid construct of claim 1 , wherein the therapeutic agent is selected from the group consisting of an anti-cancer agent, an apoptosis inducing agent, a tumor suppressor agent, an immune system enhancing agent, and an immunomodulatory agent.

4. The nucleic acid construct of claim 1 , wherein the therapeutic agent is mda-7/IL-24.

5. An isolated mammalian cell comprising the nucleic acid construct of claim 1 .

6. The mammalian cell of claim 5 , wherein the cell is a cancer cell.

7. A viral vector comprising the nucleic acid construct of claim 1 , wherein the viral vector is selected from the group consisting of an adenoviral vector, a lentiviral vector, a retroviral vector, an adeno-associated viral vector, and a herpes simplex viral vector.

8. A nanoparticle comprising the nucleic acid construct of claim 1 .

9. A method of imaging and treating cancerous cells in a subject in need thereof, comprising the steps of: administering to the subject a nucleic acid construct of claim 1 , wherein the gene encoding the avidin or streptavidin and the gene encoding the therapeutic agent are expressed in cancerous cells of the subject; administering to the subject a biotinylated adduct comprising an imaging agent; and detecting a signal from the imaging agent.

10. The method of claim 9 , wherein the cancerous cells are selected from the group consisting of breast cancer, melanoma, carcinoma of unknown primary (CUP), neuroblastoma, malignant glioma, cervical, colon, hepatocarcinoma, ovarian, lung, pancreatic, and prostate cancer.

11. The method of claim 9 , wherein a viral vector comprises the nucleic acid construct, and wherein the viral vector is selected from the group consisting of an adenoviral vector, a lentiviral vector, a retroviral vector, an adeno-associated viral vector, and a herpes simplex viral vector.

12. The method of claim 9 , wherein a nanoparticle comprises the nucleic acid construct.

13. The method of claim 9 , wherein the second promoter is a cancer selective promoter.

14. The method of claim 9 , wherein the therapeutic agent is selected from the group consisting of an anti-cancer agent, an apoptosis inducing agent, a tumor suppressor agent, an immune system enhancing agent, and an immunomodulatory agent.

15. The method of claim 9 , wherein the therapeutic agent is mda-7/IL-24.

16. A method of treating cancerous cells in a subject, comprising the steps of: a. administering to the subject a nucleic acid construct comprising a first promoter operably linked to a gene encoding avidin or streptavidin comprising a membrane localization signal and a transmembrane domain, and a second promoter operably linked to a gene encoding a first therapeutic agent, wherein the first promoter is cancer-selective, wherein the construct comprises a sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, and wherein the gene encoding the therapeutic agent is expressed in the cancerous cells; and b. contacting the cancerous cells with a biotinylated adduct comprising a second therapeutic agent.

17. A composition comprising an ultrasound targeted microbubble population, wherein the microbubble population stably binds a nucleic acid construct comprising a first promoter operably linked to a gene encoding avidin or streptavidin comprising a membrane localization signal and a transmembrane domain, and a second promoter operably linked to a gene encoding a therapeutic agent, wherein the first promoter is cancer-selective, and wherein the construct comprises a sequence selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18.

18. A method for delivering a nucleic acid construct to cancerous cells in a subject, comprising the steps of:
a. providing an ultrasound targeted microbubble population stably binding the nucleic acid construct of claim 1 ;
b. providing an ultrasound device capable of directing the microbubble population to the cancer cells;
c. directing the microbubble population to the cancer cells with the ultrasound device; and
d. bursting the microbubble population under conditions such that the nucleic acid construct is delivered to the cancer cells;
wherein the nucleic acid construct comprises a first promoter operably linked to a gene encoding avidin or streptavidin, and a second promoter operably linked to a gene encoding a therapeutic agent, wherein the first promoter is cancer-selective.

Assignees

• Univ Johns Hopkins
• Univ Virginia Commonwealth

Inventors

• Fisher Paul B
• Das Swadesh K
• Menezes Mitchell E
• Sarkar Devanand
• Pomper Martin G

Classifications

• G01N33/575

  for cancer · CPC title

• C12N2800/22

  Vectors comprising a coding region that has been codon optimised for expression in a respective host · CPC title

• A61K48/0075

  characterised by an aspect of the delivery route, e.g. oral, subcutaneous · CPC title

• B82Y15/00

  Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors · CPC title

• C12N2830/008

  cell type or tissue specific enhancer/promoter combination · CPC title