1. Ashley Kang
Faculty Advisor: Andreas Stahl, PhD
School of Public Health
Honors Thesis
May 16, 2016
The Effect of Coenzyme Q (CoQ) on
Beige Adipogenesis in a Human Cell
Model
2. WAT, BAT, Beige adipocytes
• White adipose tissue (WAT)
– Large lipid droplet with few mitochondria
– Major depository for energy in lipid form
• Brown adipose tissue (BAT)
– Smaller lipid droplets, rich in mitochondria
• Expression of Uncoupling protein 1 (Ucp1)
– Main source of energy expenditure via uncoupling of oxygen
consumption from ATP production
• Dissipates chemical energy to generate heat
• Beige adipocytes
– Clusters of Ucp1-expressing adipocytes (like BAT) with thermogenic
capacity that develop in WAT depots
Sell et al., 2004
3. Coenzyme Q (CoQ)
• Necessary component of
mitochondrial electron
transport chain
– Electron carrier from complex I
and II to complex III
• Longer isoprenyl side chains equate
to a more hydrophobic molecule
• CoQ plays important role in
brown adipocyte function
– Increasing CoQ levels exogenously could be therapeutically
beneficial for a variety of metabolic diseases
4. Statins
• Most effective drugs in reducing elevated low-density lipoprotein
(LDL) cholesterol
– Can be classified into two groups: hydrophilic and lipophilic
• Atorvastatin is lipophilic
• Statins and CoQ share same precursor
– Some studies suggest statins reduce endogenous production of
cholesterol and CoQ
5. Research Questions
Can a human beige adipocyte model be established by driving
preadipocytes to differentiate into more beige adipocytes?
Given that CoQ is prevalent in BAT and that it can regulate Ucp1
levels in BAT and that statins may inhibit CoQ, will statins reduce
Ucp1 expression in BAT?
Subsequently, if we exogenously supplement CoQ, can it rescue
CoQ levels?
Hypothesis: We hypothesize that statin dependent decrease
in Ucp1 expression in human beige cells is caused by CoQ
deficiency, and supplementation of CoQ can rescue this
inhibitory effect.
6. Courtesy of Dr. Ching-Fang Chang, PhD
Can we establish a human beige model?
7. • Intralipid treatment = difficult to accurately measure CoQ levels
• CoQ supplementation only
increases levels at DIF
D10-14 (4CBA/Ator) and not
at Day -3-0
• Suggested a receptor-dependent
CoQ uptake in mature adipocytes
• IL - stimulate differentiation / inhibit Ucp1 expression by promoting
adipogenesis
Problems Using Intralipid (IL) Carrier
9. Conclusion
Demonstrated that Atorvastatin and 4-CBA consistently inhibited
CoQ levels and Ucp1 expression in beige adipocytes
However, CoQ levels were neither efficiently nor effectively
supplemented
Question of whether CoQ supplementation can rescue inhibitory
effect could not be answered since results were inconclusive
Further research is necessary – many therapeutic benefits of
CoQ supplementation, especially in protecting heart against
mitochondrial dysfunction (diabetic cardiomyopathy / statin-
induced myopathy)
10. Acknowledgments
The Stahl Lab
• Dr. Andreas Stahl, PhD
• Dr. Ching-Fang Chang, PhD
• Dr. Jiehan Li, PhD
• Kevin Tharp
• Michael Park
• Brittney Bivins
• Hyo Min Park
• Lacey Andrews
PH195
• Dr. Charlotte Smith, PhD
• Dr. Maureen Lahiff, PhD
Editor's Notes
Good afternoon, my name is Ashley. Today I will be presenting my project, “The Effect of Coenzyme Q (CoQ) on Beige Adipogenesis in a Human Cell Model
To begin, there are three types of adipocytes that can be identified in mammals.
Ucp1 = a long-chain fatty acid anion/proton symporter located in the inner membrane of the mitochondrion, uncouples mitochondrial respiration from adenosine triphosphate (ATP) synthesis
Ucp1 expression is a hallmark of BAT
I mostly focused on beige adipocytes.
So what is CoQ?
Statins are also known as HMG CoA inhibitors
Farnesyl PP = prcursor to both cholesterol and CoQ
Nuclei = blue // neutral lipids = green // Ucp1 expression = red
Many clusters of neutral lipids which is important in showing that adipogenesis had occurred as not all cells (identified by the presence of nuclei) showed lipid accumulation
Most importantly, Ucp1 expression (hallmark of BAT) was observed around the lipid clusters
The staining confirmed that these preadipocytes successfully differentiated into beige cells
For the purposes of clarification
Day -3-0 = preadipocytes, have not undergone adipogenesis
Day 10-14 = fully matured, having undergone differentiation
Originally used IL as a carrier system for CoQ
4CBA is known to inhibit CoQ Carrier was IL, and cells were given IL only or IL + CoQ10 supplementation
Take away point = at Day-3-0, CoQ supplementation did not increase CoQ levels in 4CBA or Atorvastatin treated cells
Day 10-14, CoQ supplementation increased CoQ levels in both 4CBA and Atorvastatin
We scrapped IL and used micelle to help CoQ uptake
Micelle increased CoQ levels in both Day -3-0 and Day 11-14 (though significantly between D11-14)
Showed that micelle was a better and more effective carrier for CoQ than IL was
PROBLEM IS: to address whether CoQ supplementation can rescue Ucp1 levels, we had inconclusive data – the high error bars due to pipette error/inaccuracy/poor RNA quality
