Gene Therapy

Aubrey ArenasMikah De Vera

Jairra Mae MadrigalejoJhon Neil BarrunYllian San Luis

Strategic Intervention Material

Good day students and teachers! We are assigned to make our Strategic Intervention Material or simply called SIM all about Gene Therapy.

What is Gene Therapy? It is a technique for correcting defective genes that are responsible for disease development. There are four approaches in gene therapy; a normal gene inserted to compensate for a nonfunctional gene; an abnormal gene traded for a normal gene; an abnormal gene repaired through selective reverse mutation; it change the regulation of gene pairs.

And as we go on to our lesson, we will learn different things that will enhance our knowledge and will help us encourage many people to cure certain diseases like hemophilia, leukemia, etc. And also we want to teach them the studies that we’ve made and to help them understand more easily from our easy module.

- Miracle Therapy

Introduction:

Directions: Use the following pictures to identify the terms about gene therapy.

A G E R E P

S E T H S A

P R O N K I

The basic unit of heredity Proteins carry out most of life’s function. When altered causes dysfunction of a protein

A L C R E V

S E T L S A

V R O N K I

It is the basic structural, functional and biological unit of all known living organism.

The smallest unit of life that is classified as a living thing, and are often called the "building blocks of life".

4 pics, 1 word

G G C. E E .II E T R E NN N E N G I

Also called genetic modification, is the direct manipulation of an organism's genome using biotech.

A L U R E V

S E K L S A

V R O M K I

A malignant progressive disease in which the bone marrow and other blood-forming organs produce increased numbers of immature or abnormal leucocytes. These suppress the production of normal blood cells, leading to anemia and other symptoms.

4 pics, 1 word

4 pics, 1 word

O V C. E E .IT L R R E SC A V N G I

A tool commonly used by molecular biologists to deliver genetic material into cells. This process can be performed inside a living organism or in cell culture

R T U R E V

S E K L S A

S R O M K I

Created double stranded DNA copies from RNA genome

4 pics, 1 word

O V C. E E .IT D R R E SC A V N U I

Are double stranded DNA genome that cause respiratory, intestinal, and eye infections in humans

The inserted DNA is not incorporate into genome

R T U R E V

S E K L C A

S R O R R I

A person who has a recessive mutated gene, together with its normal allele. Carriers do not usually develop disease but can pass the mutated gene on to their children.

4 pics, 1 word

O V C. E E .IT L R R E SC A M N Y Z

A protein that facilitates a specific chemical reaction.

R T U R E V

S E K L S A

S C O N K I

 A group of identical genes, cells, or organisms derived from a single ancestor.

Pre-Test1. When was gene therapy first captulized?

a.1972 c. 1990b.1987 d. 1980

 2. It is the repair of a gene with a mutation associated with a progressive disease, prior to the expression of a medical condition, to prevent that expression?

a. Processing gene therapy c. Preventive gene therapyb. Autosomal gene therapy d. Multiple gene therapy

3. It is a type of gene therapy where the therapeutic genes are transferred into the somatic cells (non sex-cells), or body, of a patient?

a. Germ line gene therapy c. Autosomal gene therapyb. Preventive gene therapy d. Somatic gene therapy

4. These are the diseases that can cure by gene therapy except one.

a. Parkinson's disease c. Multiple myelomab. Diabetes d. Leber's congenital amaurosis

5. Who first approved gene therapy?

a. FDA c. WHOb. ADA-SCID d. DFA

Pre-Test6. When was the first gene therapy performed?

a. September 14, 1902 c. September 14, 1995b. September 14,1990 d. September 14, 1972

7. Who was the first person that was treated by gene therapy?

a. Jesse Gelsinger c. Ashanti DeSilvab. Ashenti DeSilva d. Jesse Gilsinger

8. These are the problems encountered in gene therapy except one.

a. Short life span c. Multigene Disordersb. Viral Vectors d. Fever

9. It replaces a faulty gene or adds a new gene in an attempt to cure disease or improve your body's ability to fight disease?

a. Miracles c. Herbalb. Gene therapy d. Medicines

 10. What do you mean by SCID?

a. Severe Combined Immunodeficiency c. Severe Common Immunodeficiency

b.Surgical Combined Immunodeficiency d. Stone Cloning Immunodeficiency

Miracle Therapy: Gene Are carried on a chromosome

The basic unit of heredity

Encode how to make a protein DNARNA proteins

Proteins carry out most of life’s function.

When altered causes dysfunction of a protein

When there is a mutation in the gene, then it will change the codon, which will change which amino acid is called for which will change the conformation of the protein which will change the function of the protein. Genetic disorders result from mutations in the genome.

What is Gene Therapy?It is a technique for correcting defective genes that are responsible for disease developmentThere are four approaches:

1. A normal gene inserted to compensate for a nonfunctional gene.2. An abnormal gene traded for a normal gene3. An abnormal gene repaired through selective reverse mutation4. Change the regulation of gene pairs

Gene TherapyThe Beginning…In the 1980s, Scientists began to look into gene therapy.

They would insert human genes into a bacteria cell. Then the bacteria cell would transcribe and translate the information into a

protein Then they would introduce the protein into human cellsThe First CaseThe first gene therapy was performed on September 14th, 1990

Ashanti DeSilva was treated for SCID Severe combined immunodeficiency

Doctors removed her white blood cells, inserted the missing gene into the WBC, and then put them back into her blood stream.

This strengthened her immune system Only worked for a few months

How it works?A vector delivers the therapeutic gene into a patient’s target cellThe target cells become infected with the viral vectorThe vector’s genetic material is inserted into the target cellFunctional proteins are created from the therapeutic gene causing the cell to return to a normal state

VirusesReplicate by inserting their DNA into a host cellGene therapy can use this to insert genes that encode for a desired protein to create the desired traitFour different types

Gene Therapy

RetrovirusesCreated double stranded DNA copies from RNA genome

The retrovirus goes through reverse transcription using reverse transcriptase and RNA

the double stranded viral genome integrates into the human genome using integrase

integrase inserts the gene anywhere because it has no specific site May cause insertional mutagenesis

One gene disrupts another gene’s code (disrupted cell division causes cancer from uncontrolled cell division)

vectors used are derived from the human immunodeficiency virus (HIV) and are being evaluated for safetyAdenovirusesAre double stranded DNA genome that cause respiratory, intestinal, and eye infections in humansThe inserted DNA is not incorporate into genome Not replicated though

Has to be reinserted when more cells divideEx. Common cold

Adeno-associated VirusesAdeno-associated Virus- small, single stranded DNA that insert genetic material at a specific point on chromosome 19From parvovirus family- causes no known disease and doesn't trigger patient immune response. Low information capacitygene is always "on" so the protein is always being expressed, possibly even in instances when it isn't needed.hemophilia treatments, for example, a gene-carrying vector could be injected into a muscle, prompting the muscle cells to produce Factor IX and thus prevent bleeding.

Study by Wilson and Kathy High (University of Pennsylvania), patients have not needed Factor IX injections for more than a year

Gene Therapy

Herpes Simplex VirusesDouble stranded DNA viruses that infect neuronsNon-viral OptionsDirect introduction of therapeutic DNA

But only with certain tissue Requires a lot of DNA

Creation of artificial lipid sphere with aqueous core, liposome Carries therapeutic DNA through membrane

Chemically linking DNA to molecule that will bind to special cell receptors DNA is engulfed by cell membrane Less effective

Trying to introduce a 47th chromosome Exist alongside the 46 others Could carry a lot of information But how to get the big molecule through membranes?

Problems with Gene TherapyShort Lived

Hard to rapidly integrate therapeutic DNA into genome and rapidly dividing nature of cells prevent gene therapy from long time

Would have to have multiple rounds of therapy Immune Response

new things introduced leads to immune response increased response when a repeat offender enters

Viral Vectors patient could have toxic, immune, inflammatory response also may cause disease once inside

Multigene Disorders Heart disease, high blood pressure, Alzheimer’s, arthritis and diabetes are

hard to treat because you need to introduce more than one geneMay induce a tumor if integrated in a tumor suppressor gene because insertional mutagenesis

Current StatusFDA hasn’t approved any human gene therapy product for saleReasons:In 1999, 18-year-old Jesse Gelsinger died from multiple organ failure 4 days after treatment for omithine transcarboxylase deficiency.

Death was triggered by severe immune response to adenovirus carrier January 2003, halt to using retrovirus vectors in blood stem cells because children developed leukemia-like condition after successful treatment for X-linked severe combined immunodeficiency disease

Gene Therapy

Unsuccessful Gene therapiesJesse Gelsinger, a gene therapy patient who lacked ornithine transcarbamylase activity, died in 1999.

Within hours after doctors shot the normal OTC gene attached to a therapeutic virus into his liver, Jesse developed a high fever. His immune system began raging out of control, his blood began clotting, ammonia levels climbed, his liver hemorrhaged and a flood of white blood cells shut down his lungs.

One problem with gene therapy is that one does not have control over where the gene will be inserted into the genome. The location of a gene in the genome is of importance for the degree of expression of the gene and for the regulation of the gene (the so-called "position effect"), and thus the gene regulatory aspects are always uncertain after gene therapy

Gene Therapy

Successful Gene Therapy for Severe Combine ImmunodeficiencyInfants with severe combined immunodeficiency are unable to mount an adaptive immune response, because they have a profound deficiency of lymphocytes.

severe combined immunodeficiency is inherited as an X-linked recessive disease, which for all practical purposes affects only boys. In the other half of the patients with severe combined immunodeficiency, the inheritance is autosomal recessive — and there are several abnormalities in the immune system when the defective gene is encoded on an autosome.

Severe Combine Immunodeficiency ContinuedA previous attempt at gene therapy for immunodeficiency was successful in children with severe combined immunodeficiency due to a deficiency of adenosine deaminase. In these patients, peripheral T cells were transduced with a vector bearing the gene for adenosine deaminase. The experiment was extremely labor intensive, because mature peripheral-blood T cells were modified rather than stem cells, and the procedure therefore had to be repeated many times to achieve success.

Parkinson's Disease Cont.The gene transfer procedure utilized the AAV (adeno-associated virus) vector, a virus that has been used safely in a variety of clinical gene therapy trials, and the vehicle that will be used in all of the company's first generation products, including epilepsy and Huntington's disease. In its Parkinson's disease trial, Neurologix used its gene transfer technology.

Successful One Year Gene Therapy Trial For Parkinson's Disease Neurologix a biotech company announced that they have successfully completed its landmark Phase I trial of gene therapy for Parkinson's Disease. This was a 12 patient study with four patients in each of three dose escalating cohorts. All procedures were performed under local anesthesia and all 12 patients were discharged from the hospital within 48 hours of the procedure, and followed for 12 months. Primary outcomes of the study design, safety and tolerability, were successfully met. There were no adverse events reported relating to the treatment.

Recent DevelopmentsGenes get into brain using liposomes coated in polymer call polyethylene glycol

potential for treating Parkinson’s diseaseRNA interference or gene silencing to treat Huntington’s

siRNAs used to degrade RNA of particular sequence abnormal protein wont be produced

Create tiny liposomes that can carry therapeutic DNA through pores of nuclear membrane Sickle cell successfully treated in mice

Gene Therapy

http://www.wellesley.edu/Biology/Courses/219/Gen_news/i3_Gene_Therapy.jpg

Post-Test1. When was gene therapy first captulized?

a.1972 c. 1990b.1987 d. 1980

 2. It is the repair of a gene with a mutation associated with a progressive disease, prior to the expression of a medical condition, to prevent that expression?

a. Processing gene therapy c. Preventive gene therapyb. Autosomal gene therapy d. Multiple gene therapy

3. It is a type of gene therapy where the therapeutic genes are transferred into the somatic cells (non sex-cells), or body, of a patient?

a. Germ line gene therapy c. Autosomal gene therapyb. Preventive gene therapy d. Somatic gene therapy

4. These are the diseases that can cure by gene therapy except one.

a. Parkinson's disease c. Multiple myelomab. Diabetes d. Leber's congenital amaurosis

5. Who does not approved gene therapy yet?

a. FDA c. WHOb. ADA-SCID d. DFA

Post-Test6. When was the first gene therapy performed?

a. September 14, 1902 c. September 14, 1995b. September 14,1990 d. September 14, 1972

7. Who was the first person that was treated by gene therapy?

a. Jesse Gelsinger c. Ashanti DeSilvab. Ashenti DeSilva d. Jesse Gilsinger

8. These are the problems encountered in gene therapy except one.

a. Short life span c. Multigene Disordersb. Viral Vectors d. Fever

9. It replaces a faulty gene or adds a new gene in an attempt to cure disease or improve your body's ability to fight disease?

a. Miracles c. Herbalb. Gene therapy d. Medicines

 10. What do you mean by SCID?

a. Severe Combined Immunodeficiency c. Severe Common Immunodeficiency

b.Surgical Combined Immunodeficiency d. Stone Cloning Immunodeficiency

Enrichment:Direction: Fill in the Blanks.

1.Gene therapy can use ____ to insert genes that encode for a desired protein to create the desired trait.2.The ____________ goes through reverse transcription using reverse transcriptase and RNA.3.____________ is from parvovirus family- causes no known disease and doesn't trigger patient immune response.4.__________ is a cancer that begins in developing blood cells in the bone marrow.5.___________ is a tool commonly used by molecular biologists to deliver genetic material into cells. This process can be performed inside a living organism or in cell culture.6.____________ is a gene therapy patient who lacked ornithine transcarbamylase activity, died in 1999. 7.The first gene therapy was performed on ___________________.8.__________________is a double stranded DNA virus that infects neurons.9. In the ____s, Scientists began to look into gene therapy.10.The basic unit of heredity is called ____.

Jumbled letters:Direction: Give the exact word of the jumbled letter to form a new word and described each..

1. ANODESVRIU2. EGNE3. LECL4. NEZYSM5. SEPHER IMSXELP SUVIRES6. SURVIES7. GEEITNC NENRININGEEG8. RAVIL SRTVCEO9. TREORSUVIESR 10. EULIAMKE

Crossword puzzle:

V V A L E U K E M I A F

M I S S F G X C V A Z G

O R R E T R O V I R U S

L N E U I C D J K L S E

E I I T S E M Y Z N E B

C E R R H L C A T N U N

U T R D Y L C S R M T C

L O A M U T A T I O N S

E R C L O N E H D G S X

S P O L K V B M G V F Z

Direction: Find the following words in the puzzle either horizontal, vertical, diagonal and even upside down.

Words to find:1. Virus2. Cell3. Carrier4. Retrovirus5. Enzymes6. Leukemia7. Clone8. Molecules9. Mutations10. Protein

Answer Key:Pre-Test and Post-Test Key: 1. A2. C3. B4. B5. A6. B

7. C8. D9. B10. A

4 pics, 1 word Key: 1. Gene Therapy2. Cell3. Genetic Engineering4. Leukemia5. Viral Vectors6. Retrovirus7. Adenoviruses8. Carrier9. Enzymes10. Clone

Enrichment Key: 1. Viruses2. Retrovirus3. Adeno-associated Viruses4. Leukemia5. Viral Vectors6. Jesse Gelsinger7. September 14th, 19908. Herpes Simplex Viruses9. 198010. Gene

Answer Key:Jumbled Letters: Any of the answer below.Adenovirus- is double stranded DNA genome that cause respiratory, intestinal, and eye infections in humans. The inserted DNA is not incorporate into genome.Gene- The basic unit of heredity. Proteins carry out most of life’s function. When altered causes dysfunction of a protein.Cell- It is the basic structural, functional and biological unit of all known living organism. The smallest unit of life that is classified as a living thing, and are often called the "building blocks of life".Enzymes- A protein that facilitates a specific chemical reaction.Herpes Simplex Virus- Double stranded DNA viruses that infect neuronsVirus- Replicate by inserting their DNA into a host cell. Gene therapy can use this to insert genes that encode for a desired protein to create the desired trait.Genetic Engineering- Also called genetic modification is the direct manipulation of an organism's genome using biotech.Viral Vectors- A tool commonly used by molecular biologists to deliver genetic material into cells. This process can be performed inside a living organism or in cell culture.

Retrovirus-Created double stranded DNA copies from RNA genomeLeukemia- A malignant progressive disease in which the bone marrow

and other blood-forming organs produce increased numbers of immature or abnormal leucocytes. These suppress the production of normal blood

cells, leading to anemia and other symptoms.

Answer Key:Crossword Puzzle:

As we start this project, we have all the courage we need, to do our best for this. And we give thanks to everyone behind our SIM. First and foremost we want to thank Ma’am Lim, our Biotechnology teacher for giving this project to us and letting us work as a group. Choz. Then to our parents who are helping us financially to fulfill this project especially to Aubrey’s ever loving and supportive mother for binding all the papers. To the internet who never fails to sustain our needs about gene therapy and to make it understandable and clear. Choz. For the members composing the group, Aubrey, Jairra, Mikah, Neil and Yllian. We couldn’t finish this without all our cooperation. And lastly, to our glorious and merciful God for giving us the allotted time to work on this project and giving us the wisdom to manage and think of ideas for this SIM. We’re hoping that you’ll enjoy and l earn gene therapy clearly. —Miracle Therapists

Acknowledgement:

Bases: Burdette, Walter J. The Basis for Gene Therapy. Springfield: Charles

C Thomas, 2001. Crayton, Stephanie. “First Clinical Trial Of Gene Therapy For Muscular

Dystrophy Now Under Way.” Medical News Today. 1 April 2006. University of North Carolina at Chapel Hill. 11 November 2006 <www.medicalnewstoday.com>.

Gene Therapy. Human Genome Project Information. 18 November 2005. U.S. Department of Energy Office of Science, Office

of Biological and Environmental Research, Human Genome Program. 12 September 2006 <http://www.ornl.gov/hgmis>.

McCormack, Matthew P. “Activation of the T-Cell Oncogene LMO2 after Gene Therapy for X-Linked Severe Combined

Immunodeficiency.” The New England Journal of Medicine. http://content.nejm.org. 346: 1185-1193, Apr 18, 2002.

Peel, David. “Virus Vectors & Gene Therapy: Problems, Promises & Prospects.” Virus Vectors & Gene Therapy. 1998. Department of Microbiology & Immunology, University of Leicester. 11 November 2006 <http://www.tulane.edu/

~dmsander/WWW/335/peel/peel2.html>.