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Gene Therapy and Antisense Oligonucleotides for Skin Diseases

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1. MicroRNAs are small non-coding RNA molecules that regulate gene expression at the post-transcriptional level. They play important roles in skin biology and diseases. 2. Specific microRNAs like miR-203, miR-146a and miR-125b are involved in the pathogenesis of psoriasis by regulating inflammatory signaling pathways. 3. MicroRNAs also modulate processes like wound healing, angiogenesis, and skin cancer development by targeting genes involved in these processes. Dysregulation of microRNAs has been observed in melanoma and other skin malignancies.

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GENE THERAPY M YOUSRY ABDEL-MAWLA,MD ZAGAZIG FACULTY of MEDICINE,EGYPT
DNA replication ,[object Object],[object Object]
DNA repair system: ,[object Object],[object Object]
DNA replication ,[object Object],[object Object]
Gene therapy ,[object Object],[object Object],[object Object],[object Object],[object Object]
Gene Delivery: ,[object Object],[object Object]
Ideal in vivo gene delivery system ,[object Object]
Ex vivo delivery ,[object Object]
Gene delivery systems: ,[object Object],[object Object],[object Object]
Biologic viral vectors. -Short-term expression; spreading of the infection to surrounding cell populations; new engineered vectors are avirulent in surrounding terminally differentiated cells; immunogenic. - Transduction of neurons and glial cells, wide host range; large insert size up to 30 kb; efficient infection. No ,[object Object],-Limited transduction efficiency that depends on helper viral functions, although in newer systems helper virus not needed, low efficiency of integration to genome; small insert size~4-5 kb. -Transduction of dividing and non dividing cells, all viral coding sequences can be deleted except those required for transduction; non immunogenic and nonpathogenic; long-term expression of transgene; specific integration site (some forms). Yes ,[object Object],-Expression of viral proteins results in toxic reaction and inflammation; carcinogenic; low efficiency in dividing cells, short-term expression, insert size only 7-11 kb. -Transduction of non dividing cells with high efficiency; wide host rang; high viral titer and high expression levels; newly developed gutless vectors have insert size as large as 30 kb. No ,[object Object],-Wide host range; stable transduction of dividing and non dividing terminally differentiated cells with long term expression; nonpathogenic; lack of expression of viral proteins. Yes ,[object Object],-Does not infect non dividing terminally differentiated cells, may be oncogenic optimal insert size 5-7 kb. -Wide host range; high efficiency; transduction of dividing cells; efficient expression of foreign gene product; stable integration; infects only once and does not replicate in vivo. Yes ,[object Object],[object Object],Limitations Advantages Integration To gemone Type of vectors
Non viral vectors. - Still in developmental stages. - Stable, non-infectious, can carry large fragments of DNA, non immunogenic, no integration into the genome. 4-Biologic non viral vectors human artificial chromosomes ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],- Easy to use, safe, many cell type and different applications. - Cell receptor independent, delivers genes to different tissues. - used in vaccine protocols, effective in localized area, increase gene expression, painless, no bleeding. 3- Physical methods a- Electroporation b-Gene gun approa-ch c-Microneedle injection - Random integration, inefficient DNA transfer. -- -- Unstable, remain episomal, poor gene expression. -Unstable, remain episomal, poor or gene expression. - Easy to use. - Non infectious, non immunogenic, effective for in vivo gene transfer, can carry large DNA fragments. - Targeted delivery, can carry large fragments of DNA. 2- Chemical vectors a-Calcium phosphate b-Cationic liposomes (lipoplex) c-Polylysine-DNA complexes - Transient gene expression, DNA not integrated into the genome, remain episomal. - simple, relatively efficient, non immunogenic, no mutagenesis. 1-Nacked plasmid DNA Disadvantages Advantages Vector system
Applications of gene therapy ,[object Object],[object Object],[object Object]
Applications of gene therapy ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
 
Antisense Gene Therapy (AS-ODNs)
Antisense Gene Therapy (AS-ODNs) ,[object Object]
Antisense therapy ,[object Object]
Antisense Therapy ,[object Object]
Small interfering RNA ( siRNA) ,[object Object]
Short interfering RNA( siRNA ) ,[object Object],[object Object],[object Object]
 
Antisense oligonucleotoids(AS-ODNs) ,[object Object],[object Object]
 
 
 
Methods for overcoming the skin barrier against gene delivery ,[object Object],[object Object]
MicroRNAs and the skin ,[object Object],[object Object],[object Object],[object Object]
Non-coding RNAs (ncRNA) ,[object Object],[object Object],[object Object]
Micro RNAs( MiRNAs )vs siRNAs ,[object Object],[object Object],[object Object]
MicroRNAs (miRNAs) Functions ,[object Object],[object Object]
 
The Biogenesis of miRNAs ,[object Object],[object Object],[object Object]
 
Molecular biology ,[object Object],[object Object]
Molecular biology ,[object Object]
Molecular biology ,[object Object],[object Object],[object Object]
 
MicroRNAs in Cutaneous Biology.
MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) ,[object Object],[object Object],[object Object]
 
miRNAs and psoriasis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object]
SOCS-3 Vs STAT3 ,[object Object],[object Object]
miR-146a and psoriasis ,[object Object]
miRNAs and wound healing ,[object Object],[object Object],[object Object],[object Object]
miRNAs and wound healing ,[object Object],[object Object],[object Object]
miRNAs and angiogenesis ,[object Object],[object Object]
miRNAs and skin cancer ,[object Object],[object Object],[object Object],[object Object],[object Object]
miR and Melanoma ,[object Object],[object Object],[object Object],[object Object],[object Object]
miR-221 & miR-222 in Melanoma ,[object Object]
miR &Kaposi’s sarcoma ,[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object]
 

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