APLIKASI BIOTEKNOLOGI dalam APLIKASI BIOTEKNOLOGI dalam BIDANG KESEHATAN & FARMASIBIDANG KESEHATAN & FARMASI

Dr. rer. nat. Kartika SenjariniDr. rer. nat. Kartika Senjarini

PendahuluanPendahuluan

Applikasi Bioteknologi berbasis biologi Applikasi Bioteknologi berbasis biologi molekuler:molekuler:

Molecular diagnostic (Genetic Testing)Molecular diagnostic (Genetic Testing)PharmacogenomicsPharmacogenomicsPharmococeutical productsPharmococeutical productsGene TherapyGene Therapy

PendahuluanPendahuluan

Molecular diagnosticMolecular diagnostic: diagnosis suatu penyakit : diagnosis suatu penyakit berdasarkan informasi molekuler (genetik)berdasarkan informasi molekuler (genetik)

PharmacogenomicsPharmacogenomics: studi yang mengaitkan : studi yang mengaitkan antara informasi genetik yang dimiliki setiap antara informasi genetik yang dimiliki setiap individu (individu (genomicsgenomics) dengan teknologi ) dengan teknologi pembuatan obat (pembuatan obat (pharmacologypharmacology))

Visinya adalah mendesain dan memproduksi Visinya adalah mendesain dan memproduksi obat obatan yang sesuai dengan karakteristik obat obatan yang sesuai dengan karakteristik genetik individu/organisme genetik individu/organisme

PendahuluanPendahuluan

Pharmococeutical productsPharmococeutical products: obat : obat tradisional dari tradisional dari trial and error, trial and error, Biotek Biotek memungkinkan pruduksi obat/senyawa memungkinkan pruduksi obat/senyawa biokimiawi tertentu dengan memanfaatkan biokimiawi tertentu dengan memanfaatkan infromasi genetik target dan organisme infromasi genetik target dan organisme

Gene Therapy:Gene Therapy: insersi gen pada sel atau insersi gen pada sel atau jaringan individu untuk memperbaiki jaringan individu untuk memperbaiki kerusakan/penyakit genetik khususnya kerusakan/penyakit genetik khususnya yang bersifat menurunyang bersifat menurun

Molecular diagnosticMolecular diagnostic

Berbasis Test Genetik i.e. test biokimiawi Berbasis Test Genetik i.e. test biokimiawi pada materi genetik seseorang (Human pada materi genetik seseorang (Human Genome: 25.000 – 35.000 gen)Genome: 25.000 – 35.000 gen)

Peta genetik: Peta genetik: personal anchestry, inherited personal anchestry, inherited deseasedesease

Keberadaan kelainan genKeberadaan kelainan genGenetic disorderGenetic disorder: “kemungkinan : “kemungkinan

terjadinya” dan “penurunan” kelainan terjadinya” dan “penurunan” kelainan genetikgenetik

Molecular diagnosticMolecular diagnostic

Keuntungan:Keuntungan:Menciptakan teknologi screening yang Menciptakan teknologi screening yang

baru, cepat, efisien dan spesifik (personal baru, cepat, efisien dan spesifik (personal specificity)specificity)

Basis pengembangan teknologi Basis pengembangan teknologi penyembuhan penyakit baru sekaligus penyembuhan penyakit baru sekaligus monitoring aplikasinyamonitoring aplikasinya

Prediksi respon pasien terhadap Prediksi respon pasien terhadap penanganan/penyembuhan penyakit penanganan/penyembuhan penyakit

Molecular diagnostic: PCRMolecular diagnostic: PCR

Molecular diagnostic: PCRMolecular diagnostic: PCR

Molecular diagnostic: CancerMolecular diagnostic: Cancer

Kanker: kesalahan komunikasi sel sehingga Kanker: kesalahan komunikasi sel sehingga pembelahan sel tidak dapat dikendalikanpembelahan sel tidak dapat dikendalikan

Tantangan untuk mengatasi: menentukan Tantangan untuk mengatasi: menentukan “troublemaker”-nya i.e. menentukan gen-gen “troublemaker”-nya i.e. menentukan gen-gen dan protein yang bertanggung jawab dalam dan protein yang bertanggung jawab dalam “miss-communication” tersebut“miss-communication” tersebut

Hal ini berarti: analisa biomolekul yang terdapat Hal ini berarti: analisa biomolekul yang terdapat didalam seldidalam sel

The earlier the detection and diagnosis can The earlier the detection and diagnosis can occur, the betteroccur, the better

Molecular diagnostic: CancerMolecular diagnostic: Cancer

Molecular diagnostic: CancerMolecular diagnostic: Cancer

Pengetahuan Pengetahuan genomicsgenomics (studi tentang semua (studi tentang semua gen) dan gen) dan proteomicsproteomics (studi tentang semua (studi tentang semua protein) didalam sel, melalui aplikasi teknologi protein) didalam sel, melalui aplikasi teknologi mass spectrometrymass spectrometry dan dan gene chips gene chips memungkinkan ditentukannya interaksi antar memungkinkan ditentukannya interaksi antar gen dan protein di dalam sel gen dan protein di dalam sel →→ “pola/mal “pola/mal aktivitas gen & protein” aktivitas gen & protein”

Molecular diagnosticMolecular diagnostic: memungkinkan : memungkinkan ditentukannya perubahan pola informasi dan ditentukannya perubahan pola informasi dan ekspresi gen ekspresi gen → → molecular signaturesmolecular signatures

DNA MicroarrayDNA Microarray ( (Gene ChipsGene Chips) memungkinkan deteksi ) memungkinkan deteksi ekspresi ratusan bahkan ribuan gen dalam satu waktu ekspresi ratusan bahkan ribuan gen dalam satu waktu

Gene TherapyGene Therapy

Terapi yang dilakukan terhadap suatu penyakit Terapi yang dilakukan terhadap suatu penyakit yang disebabkan oleh kerusakan pada gen yang disebabkan oleh kerusakan pada gen yang bertanggung jawab mengendalikan yang bertanggung jawab mengendalikan perkembangan penyakit tersebutperkembangan penyakit tersebut

Pendekatan:Pendekatan:1. insersi gen yang normal untuk 1. insersi gen yang normal untuk menggantikan gen yang nonfungsionalmenggantikan gen yang nonfungsional2. gen abnormal diperbaiki melalui 2. gen abnormal diperbaiki melalui selective selective reverse mutationreverse mutation3. merubah regulasi ekspresi gen3. merubah regulasi ekspresi gen

Gene TherapyGene Therapy

http://encarta.msn.com/media_461561269/Gene_Therapy.html

http://en.wikipedia.org/wiki/Gene_therapy

Gene TherapyGene Therapy

Non-viral OptionsNon-viral Options Direct introduction of therapeutic DNA Direct introduction of therapeutic DNA

But only with certain tissueBut only with certain tissue Requires a lot of DNA Requires a lot of DNA

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

Chemically linking DNA to molecule that will bind to special cell Chemically linking DNA to molecule that will bind to special cell receptors receptors

DNA is engulfed by cell membraneDNA is engulfed by cell membrane Less effective Less effective

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

Problems with Gene TherapyProblems with Gene Therapy Short Lived Short Lived

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

Would have to have multiple rounds of therapyWould have to have multiple rounds of therapy Immune ResponseImmune Response

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

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

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

diabetes are hard to treat because you need to introduce more than diabetes are hard to treat because you need to introduce more than one geneone gene

May induce a tumor if integrated in a tumor suppressor gene May induce a tumor if integrated in a tumor suppressor gene because insertional mutagenesisbecause insertional mutagenesis

Unsuccessful Gene therapiesUnsuccessful Gene therapies Jesse Gelsinger, a gene therapy patient who lacked ornithine Jesse Gelsinger, a gene therapy patient who lacked ornithine

transcarbamylase activity, died in 1999. transcarbamylase activity, died in 1999.

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

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

Successful One Year Gene Therapy Successful One Year Gene Therapy Trial For Parkinson's Disease Trial For Parkinson's Disease

Neurologix a biotech company announced that they Neurologix a biotech company announced that they have successfully completed its landmark Phase I have successfully completed its landmark Phase I trial of gene therapy for Parkinson's Disease. trial of gene therapy for Parkinson's Disease.

This was a 12 patient study with four patients in This was a 12 patient study with four patients in each of three dose escalating cohorts. All each of three dose escalating cohorts. All procedures were performed under local anesthesia procedures were performed under local anesthesia and all 12 patients were discharged from the and all 12 patients were discharged from the hospital within 48 hours of the procedure, and hospital within 48 hours of the procedure, and followed for 12 months. Primary outcomes of the followed for 12 months. Primary outcomes of the study design, safety and tolerability, were study design, safety and tolerability, were successfully met. There were no adverse events successfully met. There were no adverse events reported relating to the treatment.reported relating to the treatment.

Parkinson's Disease Cont.Parkinson's Disease Cont.

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

Science. 2006 Oct 6;314(5796):126-9.

‘Cancer regression in patients after transfer of genetically engineered lymphocytes.’

Liver

Lymphnode

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