Transcript P A U L I N A N... H H M I R E C I... M E N T O R : D...

PAULINA NGUYEN
HHMI RECIPIENT
MENTOR: DR. GANESH CHERALA
LAB: OSU-COLLEGE OF PHARMACY


About 1 in every 12
babies in the United
States is born with
low birth weight
Increased risk for
serious health
problems such as
lasting disabilities
and even death.
 Stunted growth
 Higher incidence of
diseases

Poor conditions in the womb
• Greater susceptibility to diseases in adult life
(Barker’s Hypothesis)

Low protein diet (LPD)–Rodent model for LBW
• Similar to human epidemiological findings
• LPD in rodent model can be manipulated in
laboratory conditions

Altered tissue development leads to
altered development of organs such as in
kidney and liver
◦ Affect renal organic transporter functionality
◦ Affects drug transport and excretion in and out
of the body

LBW individuals will need different drug
dosages
LPD
Low Birth
Weight
Various Organs
Kidney
Liver
Heart, etc.
Drug Transport & Pharmacokinetics
 Pharmacokinetics (PK)
 Study of how drug travels
through the body
A
D
M
E
o Absorption
o Distribution
o Metabolism
o Excretion
 Drug Metabolism and
Transport
 Renal Excretion
LUMEN
Basolateral
Membrane
Transporters
• OAT
• OCT
• PGP
• PEPT
• MRP

5 Organic Cation Transporters found in rat
OCT2
models
1. OCT1
2. OCT2
OCTN1
3. OCT3
4. OCTN1
5. OCTN2
OCTN2

Hypothesis
•

Low birth weight, as a result of altered
perinatal environment, leads to permanent
alterations in the expression of renal OCT
transporters.
Specific Aims
 To quantify mRNA expression of renal OCT2,
OCTN1, and OCTN2 at various ages
 To quantify protein expression of renal OCT2 at
various ages
Study Design
Weaned onto
regular diet
Birth weights noted;
litters culled
Control
Low Protein (LP)


Measure the mRNA level of OCT2, OCTN1,
and OCTN2 expression in male and female
offspring in both LP and Control groups
using RT-PCR
Measure the protein expression of OCT2 in
male and female offspring in both LP and
Control groups using western blotting
Fold Difference (relative to control)
Day 28 mRNA levels for males and females
2
1.5
1
0.5
0
-0.5
OCT2
OCTN1
-1
-1.5
-2
-2.5
-3
-3.5
Transporters
Males
Females
OCTN2
Fold Difference (relative to control)
Day 65 mRNA levels for males and females
3
2
1
0
-1
OCT2
OCTN1
OCTN2
-2
-3
-4
-5
Transporters
Males
Females
Fold difference (relative to control)
Day 150 mRNA levels for males and females
60
*
50
40
30
20
*
10
0
-10
OCT2
OCTN1
OCTN2
-20
-30
-40
-50
*
Transporters
Males
Females
*
Statistically Significant
Fold difference in Level of OCT Expression
Fold difference (relative to control)
50
*
40
30
20
*
10
0
OCT2
OCTN1
OCTN2
OCT2
OCTN1
OCTN2
OCT2
OCTN1
OCTN2
-10
-20
Day 28
Day 65
Day 150
-30
-40
*
Transporters
Males
Females
*
Statistically Significant
Density of OCT2 Band (normalize to β-actin)
0.03
0.025
0.02
0.015
0.01
0.005
0
Day 28
Day 65
Control
LBW
Day 150
Density of OCT2 Band (normalize to β-actin)
0.3
0.25
0.2
0.15
0.1
0.05
0
-0.05
Day 28
Day65
Control
LBW
Day150
Tissue
Tissue Homogenate


27 fold decrease in expression of OCT2 in
Day 150 LP females relative to control
(regular diet) females
Effect of LP diet seen in later life


Sex hormone regulation of OCT2 a possible
reason for 27 fold difference in LP Day 150
females
Sex hormone regulation of OCT2
• Testosterone up-regulates expression of OCT2
• Estrogen down-regulates expression of OCT2

Howard Hughes Medical Institute

Cripps Funding

Cherala Lab
◦
◦
◦
◦
◦
◦
◦
Dr. Ganesh Cherala
Barent DuBois
Jacob Pearson
Dr. Shobana Ganesan
Tahir Mahmood
Shawn Mahmood
Alexandru Cioara

OSU-College of Pharmacy

Dr. Kevin Ahern



Cherala G, Shapiro BH, and D'mello AP (2006) Two low protein diets differentially affect
food consumption and reproductive performance in pregnant and lactating rats and
long-term growth in their offspring. J Nutr 136: 2827–2833
Kathleen M. Giacomini, et al. (2010) Membrane transporters in drug development Nature
Reviews Drug Discovery , 215-236
Hagenbuch, B (2010) Drug Uptake Systems in Liver and Kidney: A Historic Perspective
Clinical Pharmacology & Therapeutics 87 1, 39–47