Discovery of neuro regenerative natural products from marine algae and cyanobacteria
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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

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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

  • 53% of new injuries occur within the 16-30 age group


Goals

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


  • 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


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


Neuro 2a Cells with Neurites


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


Neurite Outgrowth Controls

Treated

Control


Choosing a Concentration

Concentration (μg/ml)

1454 Fraction

Values are % lethality


  • 52 pure compounds

  • 27 crude fractions

  • One worth mentioning…..


…Symploca

www.sitesatlas.com


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

  • 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’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

  • 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%


H-1 Peak

Baseline 1

Baseline 2


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?

0.03 μg/mL

0.003 μg/mL

0.03 μg/mL

0.003 μg/mL


…..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…

  • Further investigation of Symploca compounds

  • Continued screening of crude fractions and newly purified compounds


Acknowledgements

  • Rebecca Medina

  • Gerwick Lab

  • Kevin Ahern & HHMI


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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