Discovery of Neuroregenerative Natural Products from Marine Algae and Cyanobacteria

1
Discovery of Neuro-regenerative Natural Products
from Marine Algae and Cyanobacteria

• Amanda Cordes, Dr. Doug Goeger, Dr. William
  Gerwick

2
Background

• At present, there are an estimated 243,000 people
  living with spinal cord injuries in the United
  States
• 53 of new injuries occur within the 16-30 age
  group

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Goals

• Identify a small molecule from
  algae/cyanobacteria which exhibits activity in
  stimulating nerve regeneration

4

• Many algae compounds are similar to retinoic acid
  and eicosanoids
• A marine sponge compound, Lembehyne A, promotes
  neurite outgrowth
• Known outgrowth promoters stop cell cycle
  progression, as do many of the labs
  characterized compounds

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First Defining Some Terms

• Neuro 2a Neuroblastoma Cells A mouse cancer cell
  line deriving from neurons
• Neuron Cell with capability of transmitting
  electric signals, found in nervous system
• Neurite Long, branching outgrowth from a neuron
• Differentiate Cells mature, adopt distinctive
  functions, less likely to divide

http//cancerweb.ncl.ac.uk
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Neuro 2a Cells with Neurites
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Experimental Protocol

• Neuro 2a cells are cultured in serum supplemented
  medium
• Cells then exposed to novel marine extracts,
  observed in 24 hr increments
• Neurite outgrowth compared against untreated
  control cells and ones treated with Lactacystin
  and with 8-Br-cAMP, known outgrowth promoters

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Neurite Outgrowth Controls
Treated
Control
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Choosing a Concentration
Concentration (µg/ml)
1454 Fraction
Values are lethality
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• 52 pure compounds
• 27 crude fractions
• One worth mentioning..

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Symploca
www.sitesatlas.com
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Symploca 1454 Fractions

• 1454 A
• 30 µg/mL 0.53
• 10 µg/mL 0.27
• 1454 B
• 30 µg/mL .32
• 10 µg/mL 1.2
• 1454 C
• 30 µg/mL Toxic
• 10 µg/mL .35

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Contd

• 1454 D
• 1 µg/mL 0.55
• 0.3 µg/mL 1.5
• 1454 E
• 10 µg/mL 1.9
• 3 µg/mL .88
• 1454 F
• 3 µg/mL 4.0
• 1 µg/mL .48

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Contd

• 1454 G
• 1 µg/mL 3.1
• 0.3 µg/mL 4.0
• 1454 H
• 0.3 µg/mL 5.2
• 0.1 µg/mL 0.9
• 1454 I
• 0.3 µg/mL 3.2
• 0.1 µg/mL 1.5

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Sub fractions

• H-1
• 0.003 µg/mL 4.5
• 0.001 µg/mL 0.8
• H-2
• 0.1 µg/mL 1.1
• 0.03 µg/mL 1.0

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H-1 Peak
Baseline 1
Baseline 2
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Assay Results

• H-1 Peak
• 0.03 µg/mL 1.0
• 0.01 µg/mL 1.6
• Baseline 1
• 0.003 µg/mL 1.8
• 0.001 µg/mL 0.5
• Baseline 2
• 0.003 µg/mL 2.0
• 0.001 µg/mL 0.6

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Possible Synergisms?
0.03 µg/mL
0.003 µg/mL
0.03 µg/mL
0.003 µg/mL
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..perhaps.

• H-1 Peak and Baseline 1
• 2.3
• H-1 Peak and Baseline 2
• 4.3
• 6.5
• 4.3

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..results, conclusions, and future plans

• Further investigation of Symploca compounds
• Continued screening of crude fractions and newly
  purified compounds

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Acknowledgements

• Rebecca Medina
• Gerwick Lab
• Kevin Ahern HHMI

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References

• Blanco et al., Cell Biol. Int. 2001, 25, 909
• Fenteany et al., J. Biol. Chem.1998, 273, 8545
• Aoki et al., Biochem. Biophys. Res. Com. 2001,
  289, 558