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Discovery of Neuro-regenerative Natural Products from Marine Algae and Cyanobacteria. Amanda Cordes, Dr. Doug Goeger, Dr. William Gerwick. Background. At present, there are an estimated 243,000 people living with spinal cord injuries in the United States

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discovery of neuro regenerative natural products from marine algae and cyanobacteria

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

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

background
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
goals
Goals
  • Identify a small molecule from algae/cyanobacteria which exhibits activity in stimulating nerve regeneration
slide4
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 lab’s characterized compounds
slide5

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

experimental protocol
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
choosing a concentration
Choosing a Concentration

Concentration (μg/ml)

1454 Fraction

Values are % lethality

slide10
52 pure compounds
  • 27 crude fractions
  • One worth mentioning…..
symploca
…Symploca

www.sitesatlas.com

symploca 1454 fractions
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%
cont d
Cont’d
  • 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%
cont d1
Cont’d
  • 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%
sub fractions
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%
slide16

H-1 Peak

Baseline 1

Baseline 2

assay results
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%
possible synergisms
Possible Synergisms?

0.03 μg/mL

0.003 μg/mL

0.03 μg/mL

0.003 μg/mL

perhaps
…..perhaps….
  • H-1 Peak and Baseline 1
    • 2.3%
  • H-1 Peak and Baseline 2
    • 4.3%
    • 6.5%
    • 4.3%
results conclusions and future plans
…..results, conclusions, and future plans…
  • Further investigation of Symploca compounds
  • Continued screening of crude fractions and newly purified compounds
acknowledgements
Acknowledgements
  • Rebecca Medina
  • Gerwick Lab
  • Kevin Ahern & HHMI
references
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
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