AIR-P/ATN Advances in Autism Research & Care Webinar Series! - - PowerPoint PPT Presentation

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AIR-P/ATN Advances in Autism Research & Care Webinar Series! - - PowerPoint PPT Presentation

Oct 03, 2022 99 likes •686 views

Welcome to todays AIR-P/ATN Advances in Autism Research & Care Webinar Series! This research activity was supported by a cooperative agreement UA3 MC11054 through the U.S. Department of Health and Human Services, Health Resources and

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SLIDE 1

This research activity was supported by a cooperative agreement UA3 MC11054 through the U.S. Department of Health and Human Services, Health Resources and Services Administration, Maternal and Child Health Research Program to the Massachusetts General Hospital. This work was conducted through the Autism Speaks Autism Treatment Network serving as the Autism Intervention Research Network on Physical Health.

Welcome to today’s AIR-P/ATN Advances in Autism Research & Care Webinar Series!

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SLIDE 2

RELATIONSHIP BETWEEN GASTROINTESTINAL DISORDERS AND STRESS REACTIVITY, IMMUNITY, AND BLOOD SEROTONIN IN AUTISM SPECTRUM DISORDERS

David Beversdorf, M.D. Principal Investigator AIR-P/ATN Webinar Monday, November 24, 2014

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SLIDE 3

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder

Investigators / Collaborating Sites:

David Beversdorf, M.D.

Missouri

Jeremy Veenstra-VanderWeele, M.D.

VUMC

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SLIDE 4

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder

BACKGROUND

  • High rate of GI disorders in ASD, but biology and etiology is

unknown

  • In general population, strong relationship between stress and GI
  • There is an augmented stress response in ASD
  • Therefore, it is possible for GI symptoms in ASD to vary as a

function of stress reactivity

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SLIDE 5

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder

Pilot Study

  • Conducted at the University of Missouri Thompson Center for Autism and

Neurodevelopmental Disorders

  • 16 children(mean age 12.3±3.6 years(±sdev), range 5-19)
  • 8 children with ASD with GI problems, 8 with ASD without GI
  • GI symptomatology verified by parent report on the Questionnaire
  • n Pediatric Gastrointestinal Symptoms – Rome III Version (QPGS

Rome-III)

  • Skin Conductance data collected during rest and in response to

vibrotactile and cold pressor stimulation (stressors)

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SLIDE 6

Figure 1. Mean skin conductance (microsiemens) for ASD GI

and ASD NO GI groups during baseline. *p < .05.

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder

Pilot Data

*

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SLIDE 7

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder Figure 2. Mean skin conductance (microsiemens) for ASD GI

and ASD NO GI groups during vibrotactile stimulation (p = 0.083)

Pilot Data

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SLIDE 8

Relationship Between Gastrointestinal Disorders and Stress Reactivity, Immunity, and Blood Serotonin in Autism Spectrum Disorder

  • Next Steps
  • Explore in a more definitive manner in a larger

sample

  • Examine other systems that may be affected by GI

disorders in ASD

  • A three-aim study was proposed…
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SLIDE 9

80 Children from ATN Registry from Missouri

Aim 1 – Explore Relationship Between GI Problems and Stress

40 Children from ATN Registry from Vanderbilt ECG and GSR Data Recordings During Stress Stimuli HRV and Mean Skin Conductance Is Stress Implicated in GI Problems in ASD?

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SLIDE 10

Aim 1 – Explore Relationship Between GI Problems and Stress

  • Hypothesis
  • Heart Rate Variability as GI symptoms increase
  • Skin Conductance as GI symptoms increase
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SLIDE 11
  • Skin Conductance Level (SCL)
  • Average conductance over time
  • Heart Rate Variability (HRV)
  • pNN50 – percentage of normal R-R

interval pairs that differ by 50ms or more.

Aim 1 – Explore Relationship Between GI Problems and Stress

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SLIDE 12

80 Children from ATN Registry from Missouri

Aim 2 – Examine Immune and Endocrine Markers in GI Disorders in ASD

40 Children from ATN Registry from Vanderbilt Collect Blood and Saliva Samples Examine IL-6 and TNF-alpha and salivary cortisol

Are there immune and endocrine differences in those with GI and ASD?

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SLIDE 13

Hypothesis

  • Immunological Functioning
  • Altered IL-6 and TNF-alpha cytokine levels in those

with greater stress reactivity

  • Cortisol
  • Salivary cortisol as GI symptoms

Aim 2 – Examine Immune and Endocrine Markers in GI Disorders in ASD

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SLIDE 14

Atypical Immune Response in ASD

  • Immune markers affected in ASD: IL-12, IFN-γ, IL-2, IL-

6, IL-10, and TNF-alpha (Ashwood et al., 2006)

  • TNF-α, IL-12, IL-10, and IL-6 are associated with GI

symptomatology (Ashwood et al, 2006; Jyonouchi et al., 2011)

  • Significant overlap between what is observed in ASD

and immune markers associated with the stress response (IL-6 and TNF-α) Cortisol Levels Increased in ASD and in GI Alone

  • Increased stress response in ASD (Corbett et al., 2008)
  • In general population, increased stress response in those

with GI disorders (Lyte, et al., 2011)

Aim 2 – Examine Immune and Endocrine Markers in GI Disorders in ASD

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SLIDE 15

80 Children from ATN Registry from Missouri 40 Children from ATN Registry from Vanderbilt Collect Whole Blood Samples Examine Whole Blood Serotonin Levels Are there differences in serotonin levels in ASD and GI?

Aim 3 – Examine Whole Blood Serotonin and GI in ASD

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SLIDE 16

Gastrointestinal Disorder Assessment

  • Questionnaire on Pediatric

Gastrointestinal Symptoms, Rome III Version (QPGS-RIII)

  • Measures frequency, severity, and

duration of functional GI disorders

  • Can be used to assess whether a

person meets criteria for a GI disorder

  • We created a scoring algorithm to

assess a range of GI symptoms

  • Higher scores = greater GI impairment
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SLIDE 17

Selection of Participants at Each Site

Screened ATN Registry Patients Contacted QPGS Rome-III Questionnaire Administered Inclusion Criteria

  • ASD Diagnosis
  • Age 6-18

Exclusion Criteria

  • Mitochondrial/genetic

disorder

  • Sensitivity to

adhesives

  • Inability to remain still
  • Bleeding disorder
  • Medical Condition
  • Lack of reliable

informant

  • Inability/unwilling to

provide consent

Invited to Participate in Study

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SLIDE 18

Study Demographics

Sex Number Percentage of Sample Male 108 90 Female 12 10 Mean Age SD Min Max N 11.89 3.75 6 18 120

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SLIDE 19

Aim 1 - Results

Draft analysis -- not audited MGH Biostatistics Center Data assembled on 20141110 ATN/AIR-P -- GI/Stress Root-Arcsine Transformed pNN50 Baseline, Cold Pressor-Baseline, Vibro-Baseline on GI Sum Scores

Scatterplot With Regression Line

10 20 30 40 50

Lower GI Sum R3

  • 0.4

0.0 0.4

Vibro-Base

10 20 30 40 50

Upper GI Sum R3

  • 0.4

0.0 0.4

Vibro-Base

10 20 30 40 50

Lower GI Sum R3

  • 0.4
  • 0.2

0.0 0.2 0.4

Cold-Base

10 20 30 40 50

Upper GI Sum R3

  • 0.4
  • 0.2

0.0 0.2 0.4

Cold-Base

10 20 30 40 50

Lower GI Sum R3

0.0 0.2 0.4 0.6 0.8

Baseline

10 20 30 40 50

Upper GI Sum R3

0.0 0.2 0.4 0.6 0.8

Baseline

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SLIDE 20

Draft analysis -- not audited MGH Biostatistics Center Data assembled on 20141110 ATN/AIR-P -- GI/Stress Root-Arcsine Transformed pNN50 Baseline, Cold Pressor-Baseline, Vibro-Baseline on GI Sum Scores

Scatterplot With Regression Line

10 20 30 40 50

Lower GI Sum R3

  • 0.4

0.0 0.4

Vibro-Base

10 20 30 40 50

Upper GI Sum R3

  • 0.4

0.0 0.4

Vibro-Base

10 20 30 40 50

Lower GI Sum R3

  • 0.4
  • 0.2

0.0 0.2 0.4

Cold-Base

10 20 30 40 50

Upper GI Sum R3

  • 0.4
  • 0.2

0.0 0.2 0.4

Cold-Base

10 20 30 40 50

Lower GI Sum R3

0.0 0.2 0.4 0.6 0.8

Baseline

10 20 30 40 50

Upper GI Sum R3

0.0 0.2 0.4 0.6 0.8

Baseline Draft analysis -- not audited MGH Biostatistics Center Data assembled on 20141110 ATN/AIR-P -- GI/Stress GSR Baseline and Cold Pressor on GI Sum Scores

Scatterplot With Regression Line

10 20 30 40 50

Lower GI Sum R3

  • 6
  • 2

2 6

Cold-Base

10 20 30 40 50

Upper GI Sum R3

  • 6
  • 2

2 6

Cold-Base

10 20 30 40 50

Lower GI Sum R3

2.5 5.0 7.5 10.0 12.5

Cold Presor

10 20 30 40 50

Upper GI Sum R3

2.5 5.0 7.5 10.0 12.5

Cold Presor

10 20 30 40 50

Lower GI Sum R3

5 10 15

Baseline

10 20 30 40 50

Upper GI Sum R3

5 10 15

Baseline

Aim 1 - Results

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SLIDE 21

Summary of findings Spearman Correlation (95% CI) P-value pNN50 baseline Upper GI 0.18 (-.02, 0.36) †0.076 pNN50 baseline Lower GI 0.21 (.01, 0.39) *0.034 pNN50 vibration-base Lower GI

  • 0.18 (-.37, 0.02)

†0.072 pNN50 cold pressor-base Lower GI

  • 0.24 (-.42, -.04)

*0.018 GSR: cold pressor- base Upper GI

  • 0.21 (-.42, 0.02)

†0.070

Aim 1 - Results

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SLIDE 22

SUMMARY OF REGRESSION ANALYSIS

  • UPPER GI predominantly related to pNN50 baseline (small

contribution from heart rate)

  • LOWER GI predominantly driven by pNN50 cold pressor minus

baseline (but very closely interrelated with baseline Aim 1 - Results

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SLIDE 23
  • Aim 2 results are forthcoming…
  • IL-6 and TNF-alpha ELISAs are being analyzed at the moment

Aim 2 - Results

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SLIDE 24

Discussion

  • Psychophysical factors

– Parasympathetic markers (pNN50) appear to be related to Lower GI and to some extent Upper GI – GSR limited findings- technically challenging – Baseline and stimulated HRV closely related – Post-hoc frequency analysis underway to more closely examine sympathetic and parasympathetic components from HRV

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Discussion

– Psychophysical factors

  • GI symptoms also related to some behavioral aspects

(Vineland, ABC)

  • Cytokines completed, being analyzed
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Discussion

  • Would be of interest in future to look at other

factors, how stress genetics relate, microbiome

  • Future implications may also include impact
  • n treatment
  • May serve as a biomarker that impacts

treatment plan

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SLIDE 27

Missouri

  • David Beversdorf, M.D.
  • Brad Ferguson, M.A.
  • Jill Akers, Sr. LPN
  • Micah Mazurek, Ph.D.
  • Kaitlyn Hartnett
  • Briana Kille
  • Univ. of Missouri – Thompson Center for

Autism & Neurodevelopmental Disorders Vanderbilt University Medical Center

  • Jeremy Veenstra-VanderWeele, M.D./

Columbia University

  • Evon Lee, Ph.D.
  • Sarah Marler, M.A.
  • Vanderbilt Kennedy Center for Research on

Human Development MGH Biostats

  • Eric Macklin, Ph.D.
  • Lily Alstein, Ph.D.
  • Erin McDonnell, M.S.

Thank you!

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SLIDE 28

Jeremy Veenstra-VanderWeele, M.D. Associate Professor of Psychiatry, Pediatrics, and Pharmacology Medical Director, Treatment and Research Institute for Autism Spectrum Disorder Kennedy Center for Research on Human Development Vanderbilt Brain Institute

Hyperserotonemia in man and mouse: leveraging a biomarker to gain insights in autism

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SLIDE 29

Funding / Disclosures

  • National Institute of Mental Health
  • National Institute of Child Health and Development
  • Autism Speaks
  • American Academy of Child and Adolescent Psychiatry
  • Brain and Behavior Research Foundation (NARSAD)
  • Agency for Health Care Research and Quality
  • Health Resources and Services Administration
  • Seaside Therapeutics
  • Roche Pharmaceuticals
  • Novartis
  • Forest
  • Sunovion
  • SynapDx
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SLIDE 30
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SLIDE 31

Outline

  • Hyperserotonemia as Endophenotype
  • SERT Ala56 mouse

– Brain – Behavior – Gut

  • Serotonin and GI symptoms in ASD
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SLIDE 32

Outline

  • Hyperserotonemia as Endophenotype
  • SERT Ala56 mouse
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SLIDE 33

HYPERSEROTONEMIA IN ~25% OF CHILDREN WITH AUTISM

SCHAIN AND FREEDMAN, 1961

Hyperserotonemia in Autism: Half a Century of Mystery

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SLIDE 34

Whole blood serotonin

  • Serotonin = 5-HT
  •  whole blood 5-HT

– Schain and Freedman, 1961

  • Platelets =>99% of blood 5-HT

– 5-HT actually produced in intestinal enterochromaffin cells

– Taken up into platelets via SERT

  • Whole blood 5-HT is highly heritable

– Broad heritability ~ 0.99 – Associated with serotonin transporter gene SLC6A4 in males Weiss et al., 2005

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SLIDE 35

Genetic Linkage of Autism on Chromosome 17

Barondes,1994

Families with only affected males (189 families) All families (327 families) Families with affected females (138 families)

Sutcliffe et al AJHG, 2005

hSERT

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SLIDE 36

Multiple Rare SERT Variants Identified in Autism Probands

  • Rare amino acid variants

– Conserved amino acids – Collectively associated with autism in males

  • Rigid-compulsive behavior
  • Variants increased 5-HT uptake in

lymphoblastoid cell lines and transiently transfected HeLa cells

– Ala56  ~30%

Prasad et al, 2009

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SLIDE 37

SERT Gly56Ala

  • 3:1 transmission rate to

affected males

– Rigid-compulsive symptoms (P = 0.0085) – Sensory aversion (P = 0.0005)

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SLIDE 38

Caveats

  • Gly56Ala found in some healthy subjects
  • Amino acid variants add to complexity of

multiple functional SERT variants

– Enable studies in mouse models

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SLIDE 39

Outline

  • Hyperserotonemia as Endophenotype
  • SERT Ala56 mouse
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SLIDE 40

Normal growth. No health problems.

Construction of mSERT Gly56Ala Knock-In Mouse

loxP

X

129S4/Jae Protamine-Cre Transgenic Mouse

129S6/SvEvTac

Tammy Jessen, Brent Thompson

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SLIDE 41

Brain

  • Increased 5-HT clearance

– Increased SERT phosphorylation

  • Increased receptor sensitivity
  • Increased 5-HT neuron firing
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SLIDE 42

Biomarker

P < 0.05

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SLIDE 43

Behavior

  • Decreased sociability

– Complicated by genetic background

  • Back out in tube test
  • Decreased ultrasonic vocalizations
  • Increased climbing/hanging behavior in home

cage

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SLIDE 44

Gastrointestinal System

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SLIDE 45

Gastrointestinal System in ASD

  • Increase in constipation in autism
  • Pat Levitt and others
  • Constipation + (Diarrhea or Soiling)
  • ~ Overflow fecal incontinence / encopresis
  • Associated with parent report of repetitive behavior and

compulsivity (P’s < 0.001)

  • OCD diagnosis (P < 0.001)
  • Observed rituals on ADOS (P = 0.009)
  • DSM-IV rigid-inflexible behavior (P = 0.014)

Peters et al., JADD, 2014

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SLIDE 46

Gastrointestinal System in SERT Mutant Mice

  • Mice lacking SERT have decreased transit

time

– Intermittent watery stool

  • SERT Ala56 mice:

– Rectal prolapse in mice > 1 year – Increased intestinal transit time

Unpublished: Kara Gross Mike Gershon

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SLIDE 47

Total and late- born submucosal neurons are deficient in SERT Ala56 mice

  • Total neurons

– HuC/D

  • Late born neurons

– TH and CGRP

WT Ala56 WT Ala56 WT Ala56 WT Ala56

Unpublished: Kara Gross Mike Gershon

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SLIDE 48

So does hyperserotonemia predict constipation in ASD?

  • “GI Stress” population, as described by Dr.

Beversdorf

– 82 subjects not taking 5-HT-related medications

  • Age 11.3 +/- 4.0
  • 74 males, 8 females
  • 75 Caucasian
  • IQ 81.5 +/- 24.5 (N = 51)
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SLIDE 49

Characterizing GI Symptoms

  • 46 have no Rome III GI disorders
  • 36 have a Rome III GI disorder

– 32 meet criteria for Functional Constipation

  • Some of these also meet broader criteria
  • Next most common = Irritable Bowel Syndrome

– Can also score quantitatively  lower GI symptom score

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SLIDE 50

Characterizing whole blood serotonin levels

  • Quantitative trait

– Log transformation to normalize – Age effect (-0.02 +/- 0.01, P = 0.05)

  • Hyperserotonemia compared to historical

samples (McBride et al., 1998)

– 20 hyperserotonemic (24%) – 62 normoserotonemic (76%)

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SLIDE 51

Primary analyses

  • Categorical functional constipation v. Ø

– Hyperserotonemia vs. historical control

  • 31% vs. 22% hyperserotonemia
  • OR 1.6 [0.52-5.3], P = 0.43

– Log 5-HT corrected for age and sex

  • Effect 0.14 [-0.03-0.31], P = 0.11
  • Continous lower GI score

– Hyperserotonemia vs. historical control

  • 14.5±10.5 vs. 18.9±13.4, P = 0.13

– Log 5-HT corrected for age and sex

  • Pearson r = 0.24 [0.01-0.45], P = 0.037, Spearman <0.05
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SLIDE 52

Secondary analyses

  • Hyperserotonemia not associated with any

behavioral variables

– Self-injury – Stereotypies – Compulsivity (P = 0.08) – Tactile sensitivity – IQ

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SLIDE 53

Next questions

  • Is this a real effect?
  • What specific lower GI symptoms drive correlation?
  • Are lower GI symptoms independently associated

with behavior?

  • What comes first, the lower GI symptoms or altered

5-HT system function? – Treat and repeat

  • What accounts for hyperserotonemia?
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SLIDE 54

Tryptophan hydroxylase activity could explain altered blood 5-HT levels

  • Tph1 expression is elevated in SERT Ala56

mice

– 5-HT synthesis increases to compensate for increased SERT-mediated re-uptake?

  • TPH1 genotype is associated with whole

blood 5-HT in relatives of children with ASD (Cross et al., 2007)

Unpublished: Kara Gross, Mike Gershon

WT Ala56

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SLIDE 55

Acknowledgements

  • Sarah Marler
  • Evon Lee
  • Brittany Peters
  • Chris Muller
  • Jenny Sauer
  • Travis Kerr
  • Ran Ye
  • Tammy Jessen
  • Brent Thompson
  • Ray Johnson
  • David Beversdorf
  • Brad Ferguson
  • Erin McDonnell
  • Lily Altstein
  • Eric Macklin
  • Randy Blakely
  • Jim Sutcliffe
  • Ana Carneiro
  • Jacki Crawley
  • Randy Blakely
  • Ed Cook
  • Bennett Leventhal
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SLIDE 56

Questions?

  • From the audience for the presenters
  • From the presenters for the audience
  • What other factors might serve as biomarkers for

gastrointestinal problems in ASD for future studies?

  • What might the implications for treatment be?
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SLIDE 57

Thank you for attending today’s ATN / AIR-P Advances in Autism Research & Care Webinar Series!

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