brain-gut-microbiota interactions and intestinal...

Wendy A. Henderson, PhD, CRNP, FAANChief, Digestive Disorders UnitDivision of Intramural Research

Brain-Gut-Microbiota Interactions and Intestinal Health

Wendy A. Henderson, PhD, MSN, CRNP

Early Life Brain-Gut Microbiota Signaling

Cong, Henderson, et al., Advances in Neonatal Care, 2015


Nutri-epigenetics

Joseph, Abey, Henderson, ONF, 2016


Key Facts About Stress Effects on Intestinal Health

Up to 20% of the US population report stress induced gastrointestinal (GI) symptoms

Single most common reason for emergency visits Top 10 reasons for outpatient visits 30 billion in costs annually Evidence that chronic stress affects intestinal

health across the lifespan

Wendy A. Henderson, PhD, CRNP, FAAN

Biobehavioral Branch: Dr. Wendy Henderson

Targeted Treatments

Stress-InducedGastrointestinal

Symptoms

Phenotypic Characterization

Personalized Profile

Implementation Science


Innovation-Gastrointestinal Pain Pointer

Developed by Henderson & Zuccolotto, PHS Invention #E-175-2010

Henderson et al., Gastroenterology Nursing, 2017

Brain-Gut Natural History Study

Hypothesis:

Design:

Male/Female Age 13-45 years No known organic cause

NIH Clinical Trial #NCT00824941

STRESS

Intestinal Permeability

Microbial Translocation

Abdominal Pain

Control IBS

OverweightControl

IBS + Overweight

- +

+

-


VariableOveralln = 90

Controln = 62

IBSn = 28

IBS-Dn = 13

IBS-Cn = 12

Gender n (%)

Male 45 (50%) 33 (53.2%) 12 (42.9%) 5 (38.5%) 5 (41.7%)

Female 45 (50%) 29 (46.8%) 16 (57.1%) 8 (61.5%) 7 (58.3%)

Race n (%)

White 45 (50%) 30 (48.4%) 15 (53.6%) 8 (61.5%) 6 (50%)

Black 26 (28.9%) 17 (27.4%) 9 (32.1%) 2 (15.4%) 6 (50%)

Asian 13 (14.4%) 10 (16.1%) 3 (5.2%) 2 (15.4%) 0

Other 6 (6.7%) 5 (8.1%) 1 (3.6%) 1 (7.7%) 0

Age (yrs) 27.82 7.85 28.22 8.05 26.93 7.45 26.62 6.99 27.50 8.09

Participant CharacteristicsProtocol 09-NR-0064, PI Henderson


Non-invasive Method to Assess Intestinal Permeability

Del Valle Pinero, et al., Henderson, Clinica Chimica Acta, 2013


Novel LC-MS Method to Intestinal Permeability

Del Valle Pinero et al., Henderson, Clinica Chimica Acta, 2013


Induce Stress Experimentally

Del Valle Pinero, et al., Henderson, Clinica Chimica Acta, 2013

Protocol 09-NR-0064, PI HendersonN=126 enrolled


Human Response to Sugar Challenge and the Microbiome


Microbial taxa associated with Visceral Pain

Brain-Gut Natural History Study

NIH Clinical Trial #NCT00824941

STRESS

Intestinal Permeability


Abdominal Pain

Model of Microbial Translocation


IFABP: p = NS


IFABP: p = NS

LBP: p = 0.014


IFABP: p = NS

sCD14: p = 0.067

LBP: p = 0.014

Pathway Analysis of Significant Genes

Abey, et al., Henderson, BBA Clinical, 2016

Electron Microscopy of Gold-labeled Plasma EVs

CD9 Lysozyme

IGg2aIGg1 MUC1 MUC2

+MT Control +MT Control


Time = 0 h Time = 8 h

[-Lyz]

[+Lyz]

Top Canonical Pathways from Affymetrix

Pyridoxal 5-phosphate Salvage PathwaySalvage Pathways of Pyrimidine RibonucleotidesAtherosclerosis Signaling

Top Molecular and Cellular Functions from Affymetrix

Cell-To-Cell Signaling and Interaction Cellular Movement Cellular Assembly and Organization


0

0.5

1

1.5

2

CD9 NT5E PD-L1 IL7R CD68

Fold

RN

A or

Pro

tein

(-Ly

z / +

Lyz)

RNA [-Lyz]

RNA [+Lyz]

Protein [-Lyz]

Protein [+Lyz]

Significant RNA and Protein with Lysozyme

Nanostring analysis of epithelial cells which survived this injury for 8 h [+ vs -Lyz] shows five RNA/Proteins which are part of LYZ-specific molecular cell response to injury or stress:


Lysozyme: innate immunity and antimicrobial

Lysozyme



Mucosa Adherent Microbiome

Fourie, et al., Henderson, Gut Microbes, 2016


Chronic Stress Murine Model



Chronic Stress and Murine Microbiota

Circular cladogram (LEfSe analysis) showing taxa that are more abundantly expressed in water avoidance (WA) stressed animals (i.e. animals which exhibit persistent stress induced gastrointestinal symptoms) in green and taxa more abundantly expressed in control (C) animals in red. Enrichment of the phyla Proteobacteria (n) and Actinobacteria (c) are characteristic of WA animals.


Wendy A. Henderson, PhD, MSN, CRNPFourie et al., Henderson. Gastroenterology, 2015, 148(4):S-777.

Host-Microbiome Interaction

Effects on Epithelial Cell Barrier Function

Barrier function measured in response to Dexamethasone and Butyrate exposure:

Butyrate

Cell-cell junctions mediate gut epithelial permeability

TEER measures epithelial permeability

Robinson, Abey, et al., Henderson, under review.

ZO-1 Occludin Merge

Untreated

+DEX

Occludin: trans-membrane protein which facilitates barrierfunction of the epithelial tight junction.

ZO-1: scaffolding protein physically linking the actincytoskeleton to Occludin and other trans-membrane tightjunction proteins.

+DEX+Butyrate

Disruption of Tight Junctions by Dexamethasone and Butyrate

Robinson, Abey, et al., Henderson, under review.

Untreated ChronicDexamethasone

Targeted Treatments

Grenham et al., Frontiers in Physiology, 2011


Ongoing and Future Studies

Targeting GI Epithelial Integrity to Improve Arterial Inflammation in HIV

Collaboration with Dr. Janet Lo Assistant Professor of MedicineMassachusetts General HospitalHarvard Medical SchoolFunding: NIH 1R01HL123351

Study Aims

Aim 1: Cross-sectional study to investigate the relationship between:

Aim 2: RCT to investigate the effects of a GLP-2 analogue (teduglutide) on:

GI epithelial integrity Intestinal permeability Markers of microbial translocation, Markers of immune activation Evidence of coronary atherosclerosis/arterial

inflammation

Hypothesis

We hypothesize that increased microbial translocation resulting from impaired epithelial integrity and impaired mucosal immunitytriggers the innate immune system, causing proatherogenic monocyteand macrophage activation and atherosclerotic disease development in HIV disease.

GI Epithelial Integrity


Immune Activation

Atherosclerosis and Arterial Inflammation

Study Procedures

Gut epithelial integrity/barrier function Colonoscopy/Endoscopy

Tissue biopsyGI epithelial tight junctionsGI epithelial apoptosisCharacterize tissue T-cells and

macrophages Blood: Microbial translocation Intestinal permeability Microbiome

VISIT 3:Colonoscopy

EGD

VISIT 4:FDG-PET

VISIT 2:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 1:(Screen Visit)Cardiac CTA*

VISIT 9:Colonoscopy

EGD

VISIT 10:FDG-PET

VISIT 8:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 7:Cardiac CTA

BASELINE VISITS INTERVENTION END OF STUDY VISITS

STUDY DESIGN

60 HIV+ Subjects

10 Healthy Controls

10 HIV+ Subjects (Visits 1-2)

10 Healthy Controls (Visits 1-2)

Cross Sectional Study:

20 Healthy Controls 70 HIV+ Subjects

Flow of participants

*Only for participants who do Visits 1-4

VISIT 5 & 6(Safety Visits)

Daily SQ injection of teduglutide or

placebo for 6 months

Aim 1: 7 HIV patients and 9 Healthy Controls

VISIT 3:Colonoscopy

EGD

VISIT 4:FDG-PET




VISIT 2:Intestinal

PermeabilityDEXA

HR-pQCT


VISIT 9:Colonoscopy

EGD

VISIT 10:FDG-PET

VISIT 8:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 7:Cardiac CTA


STUDY DESIGNFlow of participants

60 HIV+ Subjects

10 Healthy Controls


10 Healthy Controls (Visits 1-2) Interventional Study:

60 HIV+ Subjects

Randomization

Aim 2: 6 HIV patients enrolledAim 1: 7 HIV patients and 9 Healthy Controls

VISIT 3:Colonoscopy

EGD

VISIT 4:FDG-PET

VISIT 2:Intestinal

PermeabilityDEXA

HR-pQCT


VISIT 9:Colonoscopy

EGD

VISIT 10:FDG-PET

VISIT 8:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 7:Cardiac CTA



60 HIV+ Subjects

10 Healthy Controls



60 HIV+ Subjects





VISIT 3:Colonoscopy

EGD

VISIT 4:FDG-PET

VISIT 2:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 1:(Screen Visit)Cardiac CTA

VISIT 9:Colonoscopy

EGD

VISIT 10:FDG-PET

VISIT 8:Intestinal

PermeabilityDEXA

HR-pQCT

VISIT 7:Cardiac CTA



60 HIV+ Subjects

10 Healthy Controls



60 HIV+ Subjects





Microbial AssaysSchematic depicting lateral flow assay in one arm

Proposed schematic for multiplexed detection of diarrhea-causing pathogens

Nonprovisional Worldwide PCT Patent Application No.: PCT/US16/32788Kim, Henderson, Abey, Fourie, & Ferguson, 2015

Microbial AssaysSchematic depicting lateral flow assay in one arm

Proposed schematic for multiplexed detection of diarrhea-causing pathogens

Worldwide PCT Patent Application No.: PCT/US16/32788Kim, Henderson, Abey, Fourie, & Ferguson, 2016


Biobehavioral Branch: Dr. Wendy Henderson

Clinical Applications

Stress-Induced Gastrointestinal

Symptoms

Gastrointestinal Pain Pointer

Personalized Profile

Implementation Science

AcknowledgmentsLab Members:Ana Diallo, PhD, MPH, BSN, Postdoctoral IRTAJeffrey Robinson, PhD, Postdoctoral IRTAChristina Boulineaux, MS, Postbac IRTAStephanie Prescott, MSN, CRNP, GPP student

Prior Lab Members:Paule Joseph, PhD, MSN, CRNP, Postdoctoral IRTA, OWD FundedKristen Weaver, PhD, BS, BSN, MSN, CRNP, GPP StudentNatneal Kenea, BS, Special VolunteerOnyi Ozyochi, BS, Special Volunteer, OTIE fundedLeeAnne Sherwin, PhD, MS, CRNP, Special VolunteerFaye Harrell, MSN, CRNP, Special VolunteerNicolaas Fourie, PhD, Special VolunteerSarah Abey, PhD, BSN, Prior- Nurse Specialist, Research

Collaborators:Kong Chen, PhD (NIDDK)Colleen Hadigan, MD (NIAID)David Kleiner, MD, PhD (NCI)Alan Remaley, MD, PhD (NHLBI, DLM)Lorenzo Leggio, MD, PhD (NIAAA & NIDA)Peter Walter, PhD, (NIDDK)

NINRDivision of Intramural ResearchDan Wang, PhDAnn Cashion, PhD, RN, FAANSue Wingate, PhD, CRNPBrian Walitt, MD, MPH

Extramural Collaborators:Margaret Heitkemper, PhD, RN, U of Washington Anthony Zuccolato, PST, Inc.Chang Hee Kim, PhD, GoDxJohn Wiley, U of MichiganGreg Goyne, Nanostring Tech. Rob Shulman, BaylorSecondGenomeNCI- Fredrick

ParticipantsCNS IRBMetabolic UnitCC Nurses & Staff

Office of Workforce DiversityOffice of Intramural Training and EducationCommunity College Summer Enrichment ProgramAmgen

Wendy A. Henderson, PhD, CRNP, FAANChief, Digestive Disorders UnitDivision of Intramural Research

Brain-Gut-Microbiota Interactions and Intestinal Health

ChronicDEX and Butyrate

Slide Number 1Early Life Brain-Gut Microbiota SignalingNutri-epigeneticsKey Facts About Stress Effects on Intestinal HealthSlide Number 5Innovation-Gastrointestinal Pain PointerBrain-Gut Natural History StudySlide Number 8Slide Number 9Slide Number 10Slide Number 11Human Response to Sugar Challenge and the MicrobiomeMicrobial taxa associated with Visceral PainBrain-Gut Natural History StudyModel of Microbial TranslocationModel of Microbial TranslocationModel of Microbial TranslocationModel of Microbial TranslocationPathway Analysis of Significant GenesElectron Microscopy of Gold-labeled Plasma EVsSlide Number 21Significant RNA and Protein with LysozymeLysozyme: innate immunity and antimicrobial Mucosa Adherent MicrobiomeChronic Stress Murine ModelChronic Stress and Murine MicrobiotaHost-Microbiome InteractionEffects on Epithelial Cell Barrier FunctionDisruption of Tight Junctions by Dexamethasone and ButyrateSlide Number 30Targeted TreatmentsOngoing and Future StudiesStudy AimsHypothesisStudy ProceduresSTUDY DESIGNSTUDY DESIGNSTUDY DESIGNSTUDY DESIGNMicrobial AssaysMicrobial AssaysSlide Number 42AcknowledgmentsSlide Number 44Slide Number 45

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