institute of psychiatry, psychology & neuroscience · department of basic and clinical...

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Sandrine Thuret, PhD

Generation of New Hippocampal Neurons in the

Adult Brain: Implication for Mental Health

INSTITUTE OF PSYCHIATRY, PSYCHOLOGY & NEUROSCIENCE

B a s i c a n d C l i n i c a lN e u r o s c i e n c e

Department of Basic and Clinical Neuroscience, Cells and Behaviour UnitInstitute of Psychiatry, Psychology and Neuroscience, King’s College London.WEBSITE https://tinyurl.com/Thuret-Lab

TWITTER @thudrine

Disclosure Statement of Financial Interest

I, (Sandrine Thuret),

DO NOT have a financial interest/arrangement or affiliation with one or

more organizations that could be perceived as a real or apparent

conflict of interest in the context of the subject of this presentation.

INSTITUTE OF PSYCHIATRY, PSYCHOLOGY & NEUROSCIENCE

B a s i c a n d C l i n i c a lN e u r o s c i e n c e

Department of Basic and Clinical Neuroscience, Cells and Behaviour UnitInstitute of Psychiatry, Psychology and Neuroscience, King’s College London.WEBSITE https://tinyurl.com/Thuret-LabTWITTER @thudrine

ADULT HIPPOCAMPAL

NEUROGENESIS

Reg

ulat

ory

Mec

hani

sms

Molecular

Environmental

Health

Disease

Prev

entio

nIn

terv

entio

n

Prediction

Mon

itorin

g

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“Once development was ended, the fonts of growth and regeneration of the axons anddendrites dried up irrevocably. In the adult centers, the nerve paths are something fixed, andimmutable: everything may die, nothing may be regenerated.”

Santiago Ramon y Cajal, 1928

Autoradiographic and histological evidence of postnatal hippocampal neurogenesis in rats.

Altman & Das, 1965

Adult Neurogenesis?

Adult hippocampal neurogenesis

N euN C A G G FP G FA P

Adult hippocampal neurogenesis in humans

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Adult hippocampal neurogenesis in humans

Functional Relevance of Adult Hippocampal Neurogenesis: Learning and Memory

New Dentate Gyrus granule cells:• Increase spatial memory capacity

• Reduce interference between memories (pattern separation)

• Add information about time to memories

• Are involved in forgetting of established context-memories.

•Adult Hippocampal Neurogenesis is reduced in some animalmodels of depression.

•Many treatments for depression promote Adult HippocampalNeurogenesis and/or are dependent on functional neurogenesis.

Functional Relevance of Adult Hippocampal Neurogenesis: Mood

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Adult Hippocampal Neurogenesis: Regulated by environmental influences

Adult Hippocampal Neurogenesis: is modifiable

controls

runners

Adult Hippocampal Neurogenesis emerging as Target of choice?

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Our hippocampal neurogenesis Lines of Research

Dem

elza

Smee

th

Amin

aM

cDia

rmid

Hyun

-ah

Lee

Chia

ra d

e Lu

cia

Curie

Kim

Thom

as

Berg

er

Andr

ea

Du P

reez

Edin

aSi

lajd

zic

Alish

Palm

os

Animal Models

=

=

=

A d L ib i t u m ( A L )

In t e r m it t e n t F a s t in g ( IF )

C a lo r ie R e s t r ic t e d ( C R )

1 0 0 %

9 0 %

9 0 %

DH VHDCX

Wt

kl/k

l

Intermittent fasting enhances recognitionmemory and adult hippocampal neurogenesisvia the longevity gene Klotho and miR-497.

Dias et al. in preparation

Understanding the molecularneurobiology of depression.

Musaelyan et al. 2018 and in preparation

CON UCMSNeurogenesis depletion using anti-mitotic drug Temozolomide causeschanges in affective behaviour inmice.

Egeland et al. Translational Psychiatry2017

Human Hippocampal Stem Cell line – Controlled Environment

+EGF+FGF

Differentiation

CellProliferationAdd

4-OHT

Remove4-OHTEGFFGF

Identification and validation of new genes and microRNAs involved

in Proliferation and/or Neuronal Differentiation

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Human Hippocampal Stem Cell line – Stress Model

Identification of drugs for

repositioning as new

antidepressants

Powel et al. 2017a, b

Identification of the mode of action of antidepressants

Anacker et al., 2011, 2013a, 2013b

Identification of nutrient-derived

bioactivespreventing stress-induced decrease of neurogenesis

Stangl et al. in preparation

AHN: A Target for intervention

HEALTHDISEASE

PREVENTION

TREATMENT

AHN: A Biomarker for Health status, disease prediction and monitoring

HEALTHDISEASE

PREDICTION

MONITORING


Human Serum

Semi-automated cell profiling platformCellular read-out: Stem cellness- Proliferation-

Differentiation (inc. neurogenesis)- Neuriteoutgrowth – Senescence- Cell death…

Aleksandra Maruszak et al., bioRxivMurphy et al. in preparation

Human Hippocampal Progenitor cell line


HEALTHDISEASE

PREDICTION

MONITORING

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Why using serum?

(i) The neurogenic niche is localized around blood vessels allowing for potential communication with the systemic environment.

(i) Cognitive/mood impairments and adult hippocampal neurogenesis can be ameliorated through systemic perturbations such as exercise and diet.

(iii) The systemic milieu can inhibit or promote adult neurogenesis in an age-dependent fashion in mice.

In vitro Parabiosis assay

Healthy Young donors(n=27, Mean age: 29.7)

Healthy Old donors(n=35, Mean age: 77.7)

Tytus Murphy et al. in preparation

Older donor serum increases apoptotic hippocampal stem cell death

Unpub

lished

data

Shown l

ive at

the m

eetin

g

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Age of Donor serum alone is not linked to hippocampal stem cell proliferation and differentiationUnp

ublish

ed da

ta

Shown l

ive at

the m

eetin

g

Chiara de Lucia, unpublished data, in preparation

Hippocampal and DG volumes are correlated with the percentage of Neuroblasts

Unpub

lished

data

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ive at

the m

eetin

g

Cognitive Decline is linked to lower percentage of Neurons

Chronological age alone does not correspond to biological age when investigating neurogenesis

Unpub

lished

data

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Does older donor serum induces a molecular ageing phenotype?

LumiR pre-processing

Functional network analysis reveals conserved ageing molecular signature

Villeda et al., 2014Murphy et al. in preparation

Data: Tytus Murphy and Chiara de LuciaImage from Lopez-Otin et al., 2013

NAMPT SOD2

DIABLO

POLGDNML1

Mitochondrialactivity

H2A.XPARP1 XRCC2

TERT

SIRT1 pSIRT1AKT

pAKT

SIRT1 pSIRT1

AMPK pAMPK

NfKBpNfKB

UCHL1

ATF4

ATG5S6 pS6

CDKN2A

CDKN2B

CDKN2D

CDKN1A

TP53

TNFR1FADD

Older donor serum induces a molecular ageing phenotype in hippocampal stem cells

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Old_Young in vitro parabiosis recapitulates some molecular hallmarks of AgeingAgeing expression profile of hippocampal stem cells exposed to Old v. Young serum

Y (Mean age: 29.7); O (Mean age: 77.7)

Ageing Cellular profile of hippocampal stem cells exposed to Old v. Young serum

Age is associated with increased heterogeneity:Reflecting a lifetime of unequal exposure to changing environments / different life styles e.g. diet à Chronological age ⍀ biological age

The systemic environment is a major determinant of hippocampal stem cell biology during ageing.

Mu & Gage, 2011Maruszak et al. 2013

Systematic comparison needed withsame:

-Age-gender-genetic background-neuropathology stage-methods of neurogenesis detection

Hippocampal neurogenesis in Alzheimer’s Disease (rodent models)

Gatt et al. 2018

Hippocampal neurogenesis in Alzheimer’s Disease (human postmortem tissues)

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Hippocampal neurogenesis in Alzheimer’s Disease

Alterations in AHN occur at the very early stage of AD progression

Prior to processes that may secondarily affect neurogenesis (neuronal loss, amyloiddeposition and inflammation).

AHN= An integral part of AD pathology

-Can we clarify the longitudinal changes in hippocampal neurogenesis during AD progression?

-Can we predict AD conversion from Mild Cognitive Impairment (MCI)?

Hippocampal neurogenesis in Alzheimer’s Disease

MCI AD

L o n g i t u d i n a l p a t i e n t s s e r u m s a m p l e s

Longitudinal serum samples AddNeuroMed and KHP-DCR cohorts

conversion

M ild cognitive impairment

Dementia due to Alzheimer’s disease

Longitudinal blood collection from MCI converters to AD(N=38, 2-6 follow-up visits)

M ild cognitive impairment (non-converters)

Longitudinal blood collection from MCI non-converters (N=18, 3-6 follow-up

visits)

Aleksandra Maruszak et al., bioRxiv 175604; doi: https://doi.org/10.1101/175604

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Can we monitor AD progression?

Signatures of disease progression

DA

PI

CC

3K

i67

MCI Alzheimer’s disease

p=0.023

p=0.002

p<0.0001

Increased cell count

Increased apoptotic cell death

Decreased proliferation

Signatures of conversion to AD (proliferation assay)

Mixed effects regression models for repeated measuresP-values represent significance of the fitted model.


Increased number of neuroblasts

MCI Alzheimer’s disease

DAP

ID

cxM

ap2

p=0.021

p=0.044

p=0.037

Increased cell count

Increased number of neurons

Signatures of conversion to AD (differentiation assay)



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Can we distinguish MCI converters from MCI non-converters?


MCI converters and non-converters have a different cellular profile

AVERAGE CELL COUNT (differentiation)

p<0.0001 p<0.0001

p=0.0001, R2= 0.099, AdjR2= 0.093

AVERAGE CELL COUNT (proliferation)

PROLIFERATION (%Ki67) APOPTOTIC CELL DEATH (%CC3, proliferation)

p=0.0001p=0.0005

NEUROGENESIS (%Map2)

p<0.0001, R2= 0.2631, AdjR2= 0.258

APOPTOTIC CELL DEATH (%CC3, differentiation)



Who will convert to AD?


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Who will convert to AD?

•Baseline serum sample data•Stepwise logistic regressionèModel predicting conversion from MCI to AD with an accuracy of 96.75%

96.75% chance of correct classification

Area under the Receiver Operating Characteristic (ROC)

curve: 0.9675 - Sensitivity cut-off: 72%

OR P>z

Education (years)

0.72 0.040

Av cell count (proliferation)

1.03 0.001

Ki67 (proliferation)

1.35 0.038

CC3 (differentiation)

3.49 0.016

Intercept 1.68e-15

0.012

p=0.0001, Pseudo R2=0.6672Hosmer-Lemeshow chi2(8) = 9.22Prob > chi2 = 0.3244


Who will convert to AD: Machine Learning Cross-validation

Area under the Receiver OperatingCharacteristic (ROC) curve: 0.93 -Sensitivity 90.3%, Specificity79.0%.


Education (years)

Av cell count (proliferation)

Ki67 (proliferation)

CC3 (differentiation)

Support Vector Machine Classifiers using theRadial-Based Kernel were trained to predictconversion status.

Performance of the classifier was assessedusing 1000 repeats of 5-fold cross-validation

Ben Liu, Alejo J Nevado-HolgadoSimon Lovestone [Oxford University, UK]

Who will convert to AD: Proteomics

SOMAScan assay on baseline serum samples of MCI converters and non-converters: 4006 different protein epitopes.

Receiver-operator characteristic-

curve for predicting conversion to

Alzheimer’s disease using a panel of

207 proteins. Area under the curve,

AUC=0.94 Sensitivity= 91.65%,

Specificity= 81.68%.Ben Liu, Alejo J Nevado-HolgadoSimon Lovestone [Oxford University, UK]


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1- Can we monitor AD progression? ✓Conversion to AD is significantly associated with changes in hippocampalprogenitor cell count, proliferation, cell death and neurogenesis.

Our human hippocampal progenitor cell-based assay enables monitoring disease progression and predicting conversion to AD

3- Can we predict AD conversion? ✓Education and baseline assay readouts on hippocampal progenitor cellcount, proliferation and cell death predict conversion to AD with highaccuracy.

2- Can we distinguish MCI converters from MCI non-converters? ✓MCI converters can be distinguished from MCI non-converters usingmarkers of proliferation, neurogenesis and cell death.

Can we modify the probability to convert from MCI to AD?

m cp (marg insconplo t) a fter S TA TA fitting log is tic regress ion m odels.A verage adjusted pred ic tion for each of the observed va lues for education or C C 3, w hile other variab les are le ft a t the ir observed va lues.

§Validation study has started.

§Up to 3.5 years for intervention to delay AD conversion/ for stratification inclinical trials.

§Assay for testing candidate molecules to rescue the conversion cellularphenotype.

§Assay for monitoring interventions/disease progression.

Our human hippocampal progenitor cell-based assay enables monitoring disease progression and predicting conversion to AD

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AcknowledgementsThomas BergerCurie KimChiara De LuciaAndrea Du PreezDemelza SmeethAmina McDiarmidHyun-ah LeeEdina SilajdzicAlish Palmos

Tytus MurphyAleksandra MaruszakMartin EgelandKsenia MusaelyanAlessandra BorsiniGisele DiasZahra HassaniDoris Stangl

Collaborators:Simon Lovestone [Oxford University, UK]Ben Liu [Oxford University, UK]Alejo J Nevado-Holgado [Oxford University, UK]

institute of psychiatry, psychology & neuroscience · department of basic and clinical...

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