成果医学遗传学11mg2014zjuof genetic disease

1、Treatment of Genetic DiseaseMing Qi 祁鸣, PhD, Dip. ABMG, FACMGHUGO LecturerCAP InspectorCenter for Genetic & Genomic MedicineJames Watson Institute of Genomic Sciences& Medical School of Zhejiang UniversityDepartment of Pathology and Lab. Med.University of Rochester Medical Center 8820-8274ZJUMS Rese

2、arch Building A713Learning ObjectivesConventional managementsSurgeryInternal medicine2. Gene therapy DiseaseInterventionSubstance or TechniqueG6PD deficiencyDrug/dietary avoidanceAntimalarial drugsPKUGalactosemia Familial hypercholesterolemia Dietary restrictionPheGalCholesterolCongenital hypothyroi

3、dismReplacement of deficient productThyroxineWilson diseaseProtein drug therapyPenicillamineSCIDReplacement of deficient enzyme/proteinBlood transfusionTreatment of Genetic Disease by Metabolic ManipulationWilson disease: Cu toxicityCharacteristics ofWilson Disease (AR) Liver disease recurrent jaund

4、ice, simple acute self-limited hepatitis-like illness, autoimmune-type hepatitis, fulminant hepatic failure, or chronic liver disease. Neurologic presentations movement disorders (tremors, poor coordination, loss of fine-motor control, chorea, choreoathetosis) or rigid dystonia (mask-like facies, ri

5、gidity, gait disturbance, pseudobulbar involvement). Psychiatric disturbance depression, neurotic behaviors, disorganization of personality, and, occasionally, intellectual deterioration. Kayser-Fleischer rings: copper deposition in Descemets membrane of the cornea and reflect a high degree of coppe

6、r storage in the body. Mild or acute hemolysis. Wilson disease: Penicillamine therapy Before/AfterGene therapyThe medical procedure involves either replacing, manipulating, or supplementing nonfunctional genes with healthy genes. In vivo and ex vivo Gene TherapyIntroduce a geneRecover the Activity o

7、f an EnzymeRecover the Activity of an Enzyme & its RegulationIntroduce a Gene & its Regulatory geneGene TherapyIn most gene therapy studies, a normal gene is inserted into the genome to replace an abnormal, disease-causing gene. A carrier molecule called a vector must be used to deliver the therapeu

8、tic gene to the patients target cells. Currently, the most common vector is a virus that has been genetically altered to carry normal human DNA. Some Types of Viruses Used as Gene Therapy Vectors: Retroviruses Adenoviruses Adeno-associated viruses (AAV) Herpes simplex viruses Liposome-mediated DNA t

9、ransferAntisense oligonucleotidesRNA Interference (RNAi)siRNA: short interfering RISC: RNA induced silencing complexRNAi: Lieberman J et al. 2003.Father of Gene Rx: Anderson WFSevere Combined Immunodeficiency Syndrome (SCID)A disease of young children because, until recently, the absence of an immun

10、e system left them prey to infections that ultimately killed them. The patient has neither cell-mediated immune responses nor is able to make antibodies.About 25% of the cases of SCID due to homozygous for defective enzyme adenosine deaminase (ADA) gene. The normal catabolism of purines is deficient

11、, toxic for T cells and B cells.Severe Combined Immunodeficiency Syndrome (SCID)adenine deaminase (ADA) deficiencyRaise the child in a strictly germfree environment: all food, water, and air to be sterilized. David, the bubble boy from Houston, survived this way until he was 12 years old. ADA defici

12、ency (SCID): Ashanti de Silva，1990 1999/9/17: Jesse Gelsinger, Arizona,the first victim of gene therapy It is but sorrow to be wise when wisdom profitsnot. -Sophocles (496?-406BC)Rosenberg LE, Schechter AN.Gene therapist, heal thyself. Science, 2000;287:1751. What factors have kept GRx from ing an e

13、ffective treatment for genetic disease? Short-lived nature of gene therapy Immune response Problems with viral vectors Multigene disorders. Long Way to Go Colour blindness corrected by gene therapy Nature, Vol. 461, No. 7261, September 16, 2009 The monkeys were trained to touch a screen when they sa

14、w coloured patches - the standard Cambridge Colour TestAll male squirrel monkeys (Saimiri sciureus), which naturally see the world in just two tones Dalton, a squirrel monkey treated with gene therapy, enjoys his new colour sense .Neitz Laboratory Lebers Congenital Amaurosis (LCA) Autosomal recessiv

15、e Early onset retinal degeneration blindness, abnormal eye movements in infancy &early childhood No treatment Many cases of LCA are caused by mutations in RPE65(Patient #3 in the CHOP-Penn Study)LCA-RPE65 Normal Bennett 6/08199320072006200520042002200120002003199919981997What it takes for The Founda

16、tion to help bring a treatment to the clinicExample: RPE65 Gene Therapy for LCA (Lancelot Trial)discover RPE65 genelink RPE65 gene to LCAAAV GT vector productionVector improvement:test in mice, dogschoose AAV2 for human triallarge scale vector productionRPE65 is found to be retinyl ester binding pro

17、teinCreate RPE65 Knockoutmouse modelDiscover RPE65 natural mutation in briard dogtreat Lancelot in 1 eyepublish Lancelot findingstreated over 50 dogs:all with improved visiontoxicity, biodistribution studiesPre-IND withFDA/CBERJacobson:Orphan drug INDapplication filedJacobson RACfor adultsHigh RACfo

18、r kidsPatient screening (phenotype, genotype)Jacobson:FDA releasePhase I holdJacobson: awaitIRB approvalHigh will applyfor FDA, IRBBegin Phase I Clinical TrialFFB has supported this project from the outset, and is still an active supporterFoundation Fighting BlindnessVisions 2008Washington, DCAugust

19、 10, 2008 Vision Recovery in Individuals with Congenital Blindness after Retinal Gene Therapy Jean Bennett, M.D., Ph.D. On behalf of the CHOP-UPenn-TIGEM-SUN TeamA collaboration with:Seconda Universit degli Studi di Napoli What are the Patients Reactions?Can I have my other eye injected? They are ha

20、ppy with the results.Bennett 6/08Leber Congenital Amaurosis (LCA)Severe retina dystrophy, in the first year of life. Visual function is usually poor and often panied by nystagmus, sluggish or near-absent pupillary responses, photophobia, high hyperopia, and keratoconus.Autosomal recessive17 genes cu

21、rrently known for LCA: GUCY2D (locus name: LCA1), RPE65 (LCA2), SPATA7 (LCA3), AIPL1 (LCA4), LCA5 (LCA5), RPGRIP1 (LCA6), CRX (LCA7), CRB1 (LCA8), CEP290 (LCA10), IMPDH1 (LCA11), RD3 (LCA12), and RDH12 (LCA13), account for, about 70% of all LCA. Hunting for New LCA GeneNicotinamide Mononucleotide Ad

22、enylyltransferase 1, NMNAT1maps to 1p36.22, near the previously mapped LCA9. not previously been linked to LCA but is associated with axonal degeneration. HomozygousNmnat1 knockout mice do not survive to birth, indicating the function of Nmnat1 is essential in mice. 4 coding exons, small 279-residue

23、 protein that has an important role in NAD+ biosynthesis, catalyzing the formation of NAD+ from nicotinamide mononucleotide (NMN) and ATP. (Falk, et al Nat Genet, 2012)Mutations identified in NMNAT1 PatientMutationCountry / Ethnicity/ Ancestry MutationCountry / Ethnicity/ Ancestry 1p.W169X:c.507GA (

24、paternal)USA / Asian (Chinese), Hispanic, Caucasian p.E257K:c.769GA (maternal)Cherokee Indian, Hispanic, Caucasian2p.W169X:c.507GA (paternal)Brazil / Turkey, Germany, Italy, Portugalp.E257K:c.769GA (maternal)Brazil / Poland, Portugal, Lebanon, Italy3p.W169X:c.507GA (maternal)Brazil / Brazilian ances

25、tryp.E257K:c.769GA (paternal)Brazil / Brazilian ancestry4p.L153V:c.457CG (paternal)p.N273D:c.817AG (paternal)Canada / Western Europeanp.E257K:c.769GA (maternal)Canada / Western European5p.V98G:c.293TG (paternal)Brazil / African and Portuguesep.E257K:c.769GA (maternal) Brazil / Hispanic Caucasian6p.V

26、151F:c.451GT (parental sample not available)Canada / Greekp.E257K:c.769GA (parental samples are not available)Canada / Greek7,8p.M35T:c.104TC (paternal)USA / Caucasianp.E257K:c.769GA (maternal)USA / Caucasian9,10p.W85X:c.255GA (maternal)Canada / British and Dutchp.E257K:c.769GA (paternal sample is not available)Canada / British and DutchMutations identified in the NMNAT1 gene of LCA Patients3-D Structure of