Method for identifying and treating cancer

Abstract

A new system for identification and treatment against cancer, specifically the mutation or deletion of an antioncogene. An ideal candidate is a patient with family history for hereditary mutations in a known antioncogene. The first method of this system identifies the mutation of a patient's at-risk antioncogene by causing a natural fluorescence only when the specific at-risk antioncogene has mutated or deleted. The second method of this system utilizes a virus to attack and dissolve cancer cells with special markers to avoid the damage to normal cells, thereby achieving the purpose of treating cancer.

Claims

1. A method for treating a cancer caused by an antioncogene, comprising the following steps: step 1) identifying the antioncogene that is at a risk for mutation, wherein a deterioration of the antioncogene causes normal cells of a patient into cancer cells; step 2) inserting at least one marker-related gene adjacent to the antioncogene in the normal cells by a gene transfection method, wherein the gene transfection method is performed immediately when the patient is determined to have the risk for the deterioration of the antioncogene, and as the antioncogene deteriorates, the at least one marker-related gene deteriorates as well; the at least one marker-related gene express a marker protein directly or express a protein for a formation of a marker protein; if the at least one marker-related gene includes more than two genes, the more than two genes are placed beside each other in order to produce the marker protein; step 3) when the marker protein is detected not to express on the cancer cells, administering a virus to attack and dissolve the cancer cells without the marker protein, wherein the normal cells are not attacked by the virus because the normal cells have the marker protein; and step 4) once the cancer cells are killed and decreased, implementing combination with chemotherapy and adding immune boosters to help flush out the virus, wherein the antioncogene is at least one gene selected from the group consisting of human BRCA1, human BRCA2, human MSH2, and human TP53, and wherein the at least one marker-related gene in step 2) is a gene expressing a protein for a formation of an actin tail, the marker protein is the actin tail, and the virus attacks the cancer cells without the actin tail.

2. The method of claim 1, wherein, the gene transfection method in step 2) is one gene transfer method selected from the group consisting of biolistic gene-gun, CaPO.sub.4, dendrimers, lipsoma, cationic polymer, electroporation, cell squeezing, sonoporation, optical transfection, protoplast fusion, impalefection, nydrodynamic delivery, magnetofection, nucleofection, and viral transduction.

3. The method of claim 1, wherein, the at least one marker-related gene includes two genes of A33R encoding a protein A33 having an amino acid sequence shown in SEQ ID NO: 1 and A36R encoding a protein A36 having an amino acid sequence shown in SEQ ID NO: 2, and the two genes are placed beside each other in order to produce the actin tail.

4. The method of claim 1, wherein, the virus in step 3) is vaccinia virus.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a flow diagram of a method for identifying a cancer caused by an antioncogene according to embodiment 1 in the present invention; and

(2) FIG. 2 is a flow diagram of a method for treating a cancer caused by an antioncogene according to embodiment 2 in the present invention.

DETAILED DESCRIPTION

(3) As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to “a protein” includes more than one protein, and reference to “a virus” includes more than one virus.

(4) It is further to be understood that use of “or” means “and/or” unless stated otherwise. Similarly, “comprise,” “comprises,” “comprising” “include,” “includes,” and “including” are interchangeable and not intended to be limiting. Also, where descriptions of various embodiments use the term “comprising,” those skilled in the art would understand that in some specific instances, an embodiment can be alternatively described using language “consisting essentially of” or “consisting of.”

(5) The foregoing general description, including the drawings, and the following detailed description are exemplary and explanatory only and are not restrictive of the present invention. The section headings used herein are for organizational purposes only and not to be construed as limiting the subject matter described.

(6) Definitions

(7) In reference to the present invention, the technical and scientific terms used in the descriptions herein will have the meanings commonly understood by one of ordinary skill in the art, unless specifically defined otherwise. Accordingly, the following terms are intended to have the following meanings:

(8) “Gene” as used herein refers to a nucleic acid region, also referred to as a transcribed region, which expresses a polynucleotide, such as an RNA. The transcribed polynucleotide can have a sequence encoding a polypeptide, such as a functional protein, which can be translated into the encoded polypeptide when placed under the control of an appropriate regulatory region. A gene may comprise several operably linked fragments, such as a promoter, a 5′ leader sequence, a coding sequence and a 3′ nontranslated sequence, such as a polyadenylation site. A chimeric or recombinant gene is a gene not normally found in nature, such as a gene in which, for example, the promoter is not associated in nature with part or all of the transcribed DNA region. “Expression of a gene” refers to the process wherein a gene is transcribed into an RNA and/or translated into a functional protein.

(9) “Gene transfer” refers to methods for introduction of recombinant or foreign DNA into host cells. The transferred DNA can remain non-integrated or preferably integrates into the genome of the host cell. Gene transfer can take place for example by transduction, using viral vectors, or by transformation of cells, using known methods, such as electroporation, protoplast fusion.

(10) “Risk gene” refers to the antioncogene that the patient has a family history of mutations of.

(11) “Therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of a vaccinia virion or pharmaceutical composition may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the vaccinia virion or pharmaceutical composition to elicit a desired response in the individual. Dosage regimens may be adjusted to provide the optimum therapeutic response. A therapeutically effective amount is also typically one in which any toxic or detrimental effects of the vaccinia virion or pharmaceutical composition are outweighed by the therapeutically beneficial effects.

(12) “Deterioration” of a gene refers to the mutation or deletion of that gene.

(13) “Skip protein” refers to the actin tail that will tell the vaccinia virus to skip that cell and instead infect a cell not presenting this skip protein.

(14) “Patient” refers to a mammal, such as a non-primate (e.g., cow, pig, horse, cat, dog, rat, etc.) or a primate (e.g., monkey or human). Preferably, the mammal is a domesticated animal, such as a dog, a cat, a mouse, a cow, a sheep, a goat, a horse, a pig, or a human subject. In some embodiments, the human is an adult patient. In some embodiments, the human is a pediatric patient.

(15) “Deterioration” or “deteriorate” as used herein refers to damage, harm, lost or destroy of a structure and function of the described gene in the present invention.

(16) “Gene transfection method” refers to a transfection method such as gene-gun, CaPO.sub.4, dendrimers, lipsoma, cationic polymer, electroporation, cell squeezing, sonoporation, optical transfection, protoplast fusion, impalefection, nydrodynamic delivery, magnetofection, nucleofection, viral transduction, etc.

(17) “Suppressor gene” refers to a gene that can express a protein to suppress a fluorescent protein expressed by a fluorescent gene.

(18) “Marker-related gene” refers to a gene that can express a marker protein directly or express a protein to effect a formation of a marker protein.

(19) The present invention provides a Preparative/Realized system which uniquely changes cancer treatment as it prepares the body genetically prior to the occurrence of cancer, and later creates an effective platform for targeting and termination of cancer cells, including a Preparative phase and a Realized phase with a co-dependent relation to each other; wherein

(20) Firstly, the Preparative phase includes a normal cell, antioncogene, a marker-related gene that can express a marker protein directly or express a protein to effect a formation of a marker protein, and a gene transfection method, wherein the marker-related gene is inserted into a genome of the normal cell containing the antioncogene by the gene transfection method in an expressive state or in a suppressive state, once the antioncogene deteriorates, the marker-related gene stops expressing the marker protein or expressing the protein to effect the formation of the marker protein if the marker-related gene is preset in the expressive state, or expresses the marker protein or expressing the protein to effect the formation of the marker protein if the marker-related gene is preset in the suppressive state. In the Preparative phase, the patient is identified to be at risk for deterioration of the known antioncogene, (such as BRCA1/2). As the deterioration of many antioncogenes have a model of genetic inheritance, this is determined by family history. Then a variety of gene transfer and modifications take place via the defined methods of transfer and modification below. These modifications prepare the patient and supervising physician for the possibility of cancer via providing easier detection and facilitating a significantly more effective treatment if the known antioncogene deteriorates.

(21) Secondly, the Realized phase comprises a detecting method, wherein, if the marker-related gene is preset in the expressive state, once the marker protein is not detected by the detecting method, the normal cell turns into the cancer cell, and the cancer cell is caused by the deterioration of the antioncogene; or, if the marker-related gene is preset in the suppressive state, once the marker protein is detected by the detecting method, the normal cell turns into the cancer cell, and the cancer cell is caused by the deterioration of the antioncogene. The Realized phase begins upon realization of the deterioration of the prepared antioncogene. Due to modifications made in the preparative phase, the physician will accurately detect that the cancer originates specifically from the deterioration of the known oncogene and no other oncogene. Then, the supervising physician will administer treatments that are only feasible if the preparative phase takes place, as they depend on the presence of certain elements in the genome which will be described further into this reading. These elements are only present if the known prooncogene deteriorates.

(22) The following embodiments will provide a detailed description on how to use the Preparative/Realized system of the present invention to identify or treat a cancer.

(23) In embodiment 1, referring to FIG. 1, a method for identifying a cancer caused by an antioncogene is provided, including step 1) identifying the antioncogene that is at a risk for mutation, wherein a deterioration of the antioncogene causes a normal cell into a cancer cell; step 2) inserting a fluorescent gene, as a marker-related gene, into a position in a genome of a patient by a gene transfection method, wherein, the fluorescent gene causes the normal cell to produce a fluorescent protein as a fluorescent marker, the position of the fluorescent gene is different from a position of the antioncogene, and step 2) is performed immediately when the patient is determined to have the risk for the deterioration of the antioncogene; step 3) inserting a suppressor gene adjacent to the antioncogene in the genome by the gene transfection method, wherein the suppressor gene suppresses the fluorescent gene; and step 4) performing a regular screen for a presence of the fluorescent marker in the patient; wherein the fluorescent marker is detected in the patient when the normal cell turns into the cancer cell, and the cancer cell is caused by the deterioration of the antioncogene.

(24) The antioncogene may be human BRCA1, human BRCA2, human MSH2, human TP53, etc.

(25) The gene transfection method in step 2) may be biolistic gene-gun, CaPO.sub.4, dendrimers, lipsoma, cationic polymer, electroporation, cell squeezing, sonoporation, optical transfection, protoplast fusion, impalefection, nydrodynamic delivery, magnetofection, nucleofection, viral transduction, etc.

(26) The fluorescent gene in step 2) may be Green Fluorescent Protein (GFP) gene.

(27) In embodiment 2, referring to FIG. 2, a method for treating a cancer caused by an antioncogene is provided, including step 1) identifying the antioncogene that is at a risk for mutation, wherein a deterioration of the antioncogene causes normal cells of a patient into cancer cells; for example, certain antioncogenes, such as BRCA1 and BRCA2, have a very strong genetic link and patients with a family history of mutations in these antioncogenes are ideal candidates; step 2) inserting at least one marker-related gene adjacent to the antioncogene in the normal cells by a gene transfection method, wherein the gene transfection method is performed immediately when the patient is determined to have the risk for the deterioration of the antioncogene, and as the antioncogene deteriorates, the at least one marker-related gene also deteriorates; the at least one marker-related gene express a marker protein directly or express a protein for a formation of a marker protein; if the at least one marker-related gene includes more than two genes, the more than two genes are placed beside each other in order to produce the marker protein; tep 3) when the marker protein is detected not to express on the cancer cells, administering a virus to attack and dissolve the cancer cells without the marker protein, wherein the normal cells are not attacked by the virus because the normal cells have the marker protein; and step 4) once the cancer cells are killed and decreased, implementing combination with chemotherapy and adding immune boosters to help flush out the virus.

(28) The antioncogene may be human BRCA1, human BRCA2, human MSH2, human TP53, etc.

(29) The gene transfection method in step 2) may be biolistic gene-gun, CaPO.sub.4, dendrimers, lipsoma, cationic polymer, electroporation, cell squeezing, sonoporation, optical transfection, protoplast fusion, impalefection, nydrodynamic delivery, magnetofection, nucleofection, viral transduction, etc.

(30) The at least one marker-related gene in step 2) may be a gene expressing a protein for a formation of an actin tail, the marker protein is the actin tail, and the virus attacks the cancer cells without the actin tail.

(31) Further, the at least one marker-related gene includes two genes of A33R encoding a protein A33 having an amino acid sequence shown in SEQ ID NO: 1 and A36R encoding a protein A36 having an amino acid sequence shown in SEQ ID NO: 2, and the two genes are placed beside each other in order to produce the actin tail.

(32) The virus in step 3) may be vaccinia virus.

(33) It should be noted that while this applies to cancer, it can be applied to other diseases caused by genetic defects as well through the same mechanism of action. The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, a person skilled in the art should understand that the present invention may be modified or equivalently substituted without departing from the spirit and scope of the present invention, which should be included within the scope of the appended claims.