Changes in Brain Volume and Cognition in Mice...

Kayode Balogun, MSc, PhD

Workshop on HIV & Women

March 3, 2018

Changes in Brain Volume and Cognition in Mice Exposed In Utero

to ABC/ 3TC-ATV/ RTV

Women and HIV

➢1.5 million HIV+ women become pregnant each year

➢Combination antiretroviral therapy (cART) recommended during pregnancy

➢Successfully prevent vertical transmission of HIV

➢Uncertainty about the potential long-term effects cART-exposed children

Insults During Fetal Growth

Adverse In utero Environment

Low Birth Weight

Preterm

Fetal Programming: Developmental Origins of Health and Disease (DOHaD)

Predisposition to chronic diseases and developmental delays

HIV & cART

HIV-Exposed Uninfected (HEU) Children

• Increased susceptibility to infection

• Stunted growth in early life

• Smaller head circumference

• Speech and language delay

•Deficits in cognitive and motor development

HEU and Cognition

Hypothesis

Perinatal cART exposure alters fetal development, and results in impaired

neurological development by adversely affecting the growing brain and nervous

system

Consideration

•Delineate the effect of the drugs from HIV

• Timely information

• Test different drug regimen

•Genetically identical—eliminates genetic variations

Objective 1

To assess the impact of in utero exposure to cART on fetal brain morphology

Approach

Female C57BL/6 mice

Pregnancy

GD1

Control

cART

GD15.5

Fetuses

Micro-CT scan➢Volumetric analysis

cART=ABC/3TC+ATV/RTV

cART was Associated with Low Fetal Weight in Pregnant c57BL/6 mice

Data = Mean and Range; Statistics = Mann Whitney; n=8 litters/group

Control cART0.0

0.2

0.4

0.6

P = 0.007F

eta

l W

eig

ht

(g)

cART was Associated with Changes in Volumetric Measurements in different

Regions of Fetal Mouse Brain

Data = Mean and Range; Statistics = Mann Whitney; n=8

Noecortex and amygdala (purple), thalamus (green),Hypothalamus (blue),

Med

ulla

Cer

ebel

lar P

rimor

dium

Neo

pallia

l cor

tex

and

amyg

dala

Hyp

otha

lam

us

Thala

mus

Pons

Spina

cor

d0.00

0.05

0.10

0.15

Control

cART

13%

10%

-7%

7%

5%

10%

8%

****

*

*

*

**

*

Norm

aliz

ed

Vo

lum

e

What about adult mice?

Approach

Female C57BL/6 mice

Pregnancy

GD1

Control

cART

WeightsMRIBehavioural tests

Delivery

cART Exposed Mice were Smaller than the Controls

Data = Median and Range; (Two way ANOVA; n= 20-22 per group)

Male Female15

20

25

30

35

ControlcARTP<0.0001

P<0.0001

We

igh

t (g

)

cART-exposed Mice had Smaller Brains than the Controls

Brain GM250

300

350

400

450

500

ControlPI-cART

Treatment: P<0.0001

Vo

lum

e (

mm

3)

Data = Median and Range; (Two way ANOVA; n= 13-18)

cART was Associated with Changes in Volumetric Measurements in Different Regions of an Adult Mice Brain (MRI)

Objective 2

To assess the impact of in utero exposure to cART on cognitive, neurosensory and

motor behaviours

cART was Associated with Changes in Volumetric Measurements in Different Regions

of an Adult Mice Brain (MRI)

Control cART0.009

0.010

0.011

0.012

0.013

0.014

P = 0.012

Se

co

nd

ary

Mo

tor

Cort

ex

No

rma

lize

d V

olu

me

Control cART0 . 0 4 5

0 . 0 5 0

0 . 0 5 5

0 . 0 6 0

0 . 0 6 5

P = 0 . 0 3

Pr

i.

S

om

at

os

en

so

ry

co

rt

ex

No

rm

al

iz

ed

V

ol

um

e

Data = Mean and Range; Statistics = Mann Whitney; n=13-16

Strength and Coordination

Joanna et al, Neuromethods, Vol. 39: Neuropeptide Techniques

Data = Median and Range; (Mann Whitney; n= 10-15 for Litter Average)

Control cART0

2

4

6

8

10

P = 0.0005

Days

The Development of Motor Skills was Delayed in Mice Exposed to cART In Utero

Tactile Reflex

The Development of Tactile Reflex was Delayed in Mice Exposed to cART In Utero

Joanna et al, Neuromethods, Vol. 39: Neuropeptide Techniques

Data = Median and Range; (Mann Whitney; n= 10-15 for Litter Average)

Control cART10

12

14

16

P = 0.0005

Days

Olfactory Reflex

The Development of Olfactory Reflex was Delayed in Mice Exposed to cART In Utero

http://www.ssponline.com/shepherds_cob.htm

Litter Averages0

20

40

60

80

ControlcART

73.8%

47.3%

P=0.0065

% P

asse

d

Data = % passed; (Fisher’s exact test; n= 10-15 for Litter Average)

Impaired cognitive function

cART was Associated with Changes in Volumetric Measurements in different Regions of an Adult

Mouse Brain (MRI)

Control cART0.014

0.015

0.016

0.017

0.018

P = 0.0016

CA

1 a

nd

CA

2N

orm

alize

d V

olu

me

Data = Mean and Range; Statistics = Mann Whitney; n=13-16

Control cART0.024

0.026

0.028

0.030

0.032

P = 0.0003

Cin

gu

late

Co

rte

x

Vo

lum

e (

mm

3)

Novel Object Recognition (NOR) Test

3hr

In utero Exposure to PI-cART is Associated with Smaller Hippocampus and a Lower Memory Index

Data = Median and Range n= 20-22; Statistics = Mann Whitney and Spearman’s correlation

Memory Index=(tn-tf)/(tn+tf)

-1.0 -0.5 0.0 0.5 1.00.024

0.026

0.028

0.030

0.032 r=0.48P=0.011

Memory Index

Cin

gu

late

Co

rte

xN

orm

alize

d V

olu

me

ControlcART

-1.0 -0.5 0.0 0.5 1.0

0.015

0.016

0.017r=0.64P=0.0003

Memory Index

CA

1 a

nd

CA

2N

orm

alize

d V

olu

me

Male Female-1.0

-0.5

0.0

0.5

1.0 P<0.0001 P<0.0001

Mem

ory

Index

Speech and Language Delay

Sociability Social Novelty

Sociability and Social Novelty Test

Male Female-1.0

-0.5

0.0

0.5

1.0

1.5

ControlcART

Pre

fere

nce Index

(So

cia

bility)

In utero Exposure to cART had no Effect on Social Interaction

Male Female-1.0

-0.5

0.0

0.5

1.0

1.5

ControlcART

Pre

fere

nce Index

(Socia

l N

ovelty)

Data = Median and Range n= 20-25; Statistics = Two-way ANOVA

Preference Index=(tn-tf)/(tn+tf)

Compared to control, in utero exposure to ABC/3TC+ATV/RTV was associated to with:

➢Lower fetal and adult weights

➢Changes in volumetric measurement in fetal and adult brains

➢Delayed motor, tactile, and olfactory reflexes

➢Lower memory index

Summary

Conclusion

Our data suggest that the in utero exposure to ABC/3TC+ ATV/RTV is associated with

volumetric changes in key regions of the brain, developmental delays, and cognitive deficits in

a mouse model of pregnancy.

Acknowledgments

• Dr. Lena Serghides

• Dr. Sharon Walmsley

• Dr. Smriti Kala

• Dr. Monica Guzman-Lenis

• Jeffrey Lu

• Pierre Lira

• Members of the Serghides’ lab

FundingOntario HIV Treatment Network Canadian Institutes of Health ResearchCanadian Foundation for AIDS ResearchOur collaborators

Collaborators• Dr. Howard Mount• Dr. John Sled• Dr. Lindsay Cahill• Lisa Yu• Jimmy Yang

www.flickr.com

Dr. Kayode Balogun is supported by a

CTN Postdoctoral Fellowship Award