Looking at Linkage Between the Processes of Autophagy and Apoptosis - Students: Zach Davis, Juliana Baumgardner, Jacob Kornilow, Joe Martin ...

Looking at Linkage Between
 the Processes of Autophagy and
            Apoptosis
Students: Zach Davis, Juliana Baumgardner, Jacob Kornilow, Joe Martin
                Research Advisor: Dr. Sherri Bergsten

Autophagy
• An intracellular process to degrade and recycle cellular material that is
  no longer needed, damaged, or harmful to the cell
   • Autophagy is accomplished through targeting of cellular components to
     autophagosomes that fuse with lysosomes

Apoptosis
• Programmed cell death which
  occurs in response to a stimulus
  that triggers a regulatory pathway

   • Intrinsic-Signal starts inside
     of the cell
   • Extrinsic-Signal starts outside
     of the cell

Methods
What conditions result in an inhibitory
versus stimulatory relationship between
autophagy and apoptosis?
•Glioblastoma Cell Line (SNB19) with
GFP-p62/DsRed-LC3 autophagic markers,
compared with vector controls
•Cells treated with drugs to modulate
autophagy and apoptosis levels
•Reverse Transcriptase qPCR used to assess
expression of regulatory genes

Results
• Reverse transcription coupled
  with qPCR provides a
  measure of RNA expression
  for each gene evaluated, by
  detection of PCR products as
  they are being made.
• Samples are done in triplicate
  to ensure accuracy.

Discussion
• Treatment with resveratrol, known to
  stimulate autophagy, also causes increased
  expression of both pro-apoptotic (BID,
  CASP10, PUMA) and anti-apoptotic (BCL-2)
  regulators.
• Inhibition of lysosomal degradation of
  autophagic cargo with chloroquine partially
  or completely blocks the stimulation of
  apoptotic regulators.
• Taken together, preliminary data suggests that
  the “toggle switch” between autophagy and
  apoptosis may occur downstream of these
  regulators, and that progression through
  autophagy is important for the impact on
  apoptosis regulation.

Future Research
• The next step is to verify the status of autophagy and apoptosis regulation
   • Studying protein levels and protein processing through Western Blots

• Further steps include trying new experimental conditions and analysis of
  other genes to compare results and determine criteria that result in
  preference for an inhibitory versus co-stimulatory relationship between the
  two processes

Acknowledgements and Literature Cited
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                                                                                                         today.