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

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

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


Background
Background Algae and Cyanobacteria

  • 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 Algae and Cyanobacteria

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



First – Defining Some Terms eicosanoids

  • 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 eicosanoids

  • 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
Neurite Outgrowth Controls eicosanoids

Treated

Control


Choosing a concentration
Choosing a Concentration eicosanoids

Concentration (μg/ml)

1454 Fraction

Values are % lethality



Symploca
…Symploca eicosanoids

www.sitesatlas.com


Symploca 1454 fractions
Symploca 1454 Fractions eicosanoids

  • 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 eicosanoids

  • 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 eicosanoids

  • 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 eicosanoids

  • 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 eicosanoids

Baseline 1

Baseline 2


Assay results
Assay Results eicosanoids

  • 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? eicosanoids

0.03 μg/mL

0.003 μg/mL

0.03 μg/mL

0.003 μg/mL


Perhaps
…..perhaps…. eicosanoids

  • 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… eicosanoids

  • Further investigation of Symploca compounds

  • Continued screening of crude fractions and newly purified compounds


Acknowledgements
Acknowledgements eicosanoids

  • Rebecca Medina

  • Gerwick Lab

  • Kevin Ahern & HHMI


References
References eicosanoids

  • 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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