Recent Advances in ALS Research

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Recent Advances in ALS Research

• At
• Solomon Park Research Institute

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Motor Neuron
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What is ALS?

• Many names
• Amyotrophic Lateral Sclerosis
• Motor Neuron Disease
• Lou Gehrigs disease.
• Motor neurons control the muscles that enable us
  to speak, eat, breathe and swallow.
• Progressive degeneration of the motor neurons in
  the brain and spinal cord.
• With no nerves to activate them, muscles
  gradually weaken and waste.
• Occurrence
• Incidence of ALS/MND is 2 per 100,000 of total
  population
• Prevalence is six per 100,000 of total population.

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• Cause is unknown
• Theories abound
• All agree that motor neurons die
• Several theories on why they die

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Research Approach at SPRI

• Our research is based on the premise that ALS is
  genetically controlled in all individuals that
  express this condition
• Goals
• Cataloguing of genes active in motor neurons
• Direct comparison of genes unique to motor
  neurons in ALS and non-ALS individuals using
  non-neural tissue (e.g. white blood cells)
• Benefits
• Will increase body of knowledge on motor neurons
• Indicate possible therapies and means of
  verifying their efficacy
• Help avoid therapies that might be harmful to
  motor neurons
• Use of non-neural tissue will circumvent
  difficulties in obtaining nervous system material
  from humans

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Introductory Genetics
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Introductory Genetics

• Genes consist of
• short coding sequences
• long non-coding sequences.
• cDNA contains uninterrupted coding sequences of a
  gene.
• The production of mRNA entails the removal of
  non-coding sequences.

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c
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Transcription
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Translation
cDNA is preferable start point when the goal or
aim is to deduce the protein sequence from DNA of
a specific tissue.
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Mouse Human Genome Similarities
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Specific Research Techniques
Technique to compare mRNA populations present in
two different tissues
Technique for amplifying specific regions of DNA
Technique for carrying desired DNA into the
bacteria for abundant synthesis of the protein
Technique to reduce the background population of
cDNAs in order to identify unique cDNAs
The process of determining exact order of
building blocks making up the DNA
Use of computers to decipher genetic code of
motor neurons
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Subtractive Hybridization
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Polymerase Chain Reaction (PCR)

• PCR Video

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Transformation
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Differential Screening
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Dot-blot Picture
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Sequencing

• The process of determining exact order of
  building blocks making up the DNA

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Motor Neuron cDNA Sequence?

• GGTACGAGCTCGGATCACTAGTAACGGCCGCCAGTGTGCTGGAATTC
  TTCGAGCGGCCGCCCGGGCAGGTCGCTGTGCGCCGCAGCACCACGGCTGG
  AGAGAAGACGGGTGTGCTGCGAAAGCGGAAGTCCTCCTCCCCGTAGAGCC
  CGGTCAGGCAGCCTCGGCACAGGCAGTAGGCTTCGGGCAGGTACCTCGGC
  CGCGACCACGCTAAGCCGAATTCTGCAGATATCCATCACACTGGCGGCCG
  CTCGAGCATGCATCTAGAGGGCCCAATTCGCCCTATAGTGAGTCGTATTA
  CAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCG
  TTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGT
  AATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCT
  GAATGGCGAATGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGT
  GGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCG
  CTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCC
  CGTCAAGCTCTAAATCGGGGGCTCCCTTTNANGGTTCCGATTTAGTGCTT
  TACGGCACCTCGACCCCAAAAACTTGATTAGGGTGATGGTTCACGTANTG
  GGCCATCGCCCTNATANACGGTTNTTCGCCCTTTTGACGTTGGAGTCACG
  TTCT

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Bioinformatics
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Identification of Complete DNA Sequence

• atgttggggacactggtctggatgctcctcgtcggcttcctgctggcact
  ggcgccgggccgcgcggcgggcgcgctgaggaccgggaggcgcccggcg
  c ggccgcgggactgcgcggaccggccagaggagctcctggagcagctgt
  ac gggcggctggcggccggcgtgctcagcgccttccaccacacgctgca
  gct cgggccgcgcgagcaggcgcgcaatgccagctgcccggccgggggc
  agggccgccgaccgccgcttccggccacccaccaacctgcgcagcgtgtc
  gccc tgggcgtacaggatttcctacgaccctgctcgctttccgaggtac
  ctgcc cgaagcctactgcctgtgccgaggctgcctgaccgggctctacg
  gggagg aggacttccgctttcgcagcacacccgtcttctctccagccgt
  ggtgctg cggcgcacagcggcctgcgcgggcggccgctctgtgtacgcc
  gaacacta catcaccatcccggtgggctgcacctgcgtgcccgagccgg
  acaagtccg cggacagtgcgaactccagcatggacaagctgctgctggg
  gcccgccgac aggcctgcggggcgctgatgccggggactgcccgccatg
  gcccagcttcc tgcatgcatcaggtcccctggccctgacaaaacccacc
  ccatgatccctg gccgctgcctaatttttccaaaaggacagctacataa
  gctttaaatatat ttttcaaagtagacactacatatctacaactatttt
  gaatagtggcagaa actattttcatattagtaatttagagcaagcatgt
  tgtttttaaacttct ttgatatacaagcacatcacacacatcccgtttt
  cctctagtaggattct tgagtgcataattgtagtgctcagatgaacttc
  cttctgctgcactgtgc cctgtccctgagtctctcctgtggcccaagct
  tactaaggtgataatgag tgctccggatctgggcacctaaggtctccag
  gtccctggagagggaggga tgtgggggggctaggaaccaagcgcccctt
  tgttctttagcttatggatg gtcttaactttataaagattaaagttttt
  ggtgttattctttc

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Specific Research Techniques
Technique to compare mRNA populations present in
two different tissues
Technique for amplifying specific regions of DNA
Technique for carrying desired DNA into the
bacteria for abundant synthesis of the protein
Technique to reduce the background population of
cDNAs in order to identify unique cDNAs
The process of determining exact order of
building blocks making up the DNA
Use of computers to decipher genetic code of
motor neurons
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Challenges

• in situ hybridization
• This will help to verify the location of the cDNA
  clones in motor neurons
• Selection of genes that would be relevant to
  motor neurons and elucidating its function
• Direct comparison of the genes of motor neurons
  between ALS and non ALS individuals

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QA