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Segregating urinary phenome responses to microbes and other factors new opportunities Chad Vezina Professor and Associate Director, George M. O’Brien Center for Benign Urology Research UW-Madison

Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

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Page 1: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Segregating urinary phenome

responses to microbes and other

factors – new opportunities

Chad Vezina

Professor and Associate Director,

George M. O’Brien Center for

Benign Urology Research

UW-Madison

Page 2: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Inflammation ꝏ progressive voiding dysfunction in men (incontinence, urinary retention, increased symptom index)Torkko et al J Urol. 2015 194:454-61

Inflammation ꝏ presence of bacteria in human prostate Hochreiter et al 2000 163:127-30

Bacterial ꝏ voiding dysfunction in mice

(↑Frequency)Lee et al PLoS One. 2015 10:e0116827

Bell-Cohn et al AJP Renal. 2019 316:F682-F692

Bacterial prostatitis ꝏ pain in mice Rudick Infect Immun. 2011 Feb;79(2):628-35

Page 3: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Histology,

Anatomy,

Microbiology

Physiology

We can see ‘Active’ inflammation /infectionbut we cannot observe past events (especially subclinical)

Standardized tests in the clinic (mostly), but what about at the bench with research animals?

Page 4: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

MAKE MOUSE VOIDING BEHAVIOR

A QUANTITATIVE TRAIT

Start with clean

mouse cageAdd filter paperAdd mouse and

wait 4 hr

Page 5: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

MAKE MOUSE VOIDING BEHAVIOR

A QUANTITATIVE TRAIT

Use Void Whizzard Software to analyze 12

features incl. spot number, size, distribution

Remove and dry

Filter paper

UV illuminate

and Image

Page 6: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Unpublished results

Wegner et al and Hill et al, Amer J Physiol-Renal 2018

Collaboration Among UW-Madison, Beaumont Health, Beth Israel / Harvard P20 Center,

Vanderbilt University

Optimized for:

• Acclimatization time

• Mouse age

• Size and shape of cage

• Assay duration

• Cage type

• Time of day

• Consistency across institutions

MAKE MOUSE VOIDING BEHAVIOR

A QUANTITATIVE TRAIT

Lisa Abler, Royal Oakes, Kyle Wegner

Page 7: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Rinse and Repeat

Human Mouse

Voiding Diary Void Spot Assay

Uroflowmetry Uroflowmetry

Cystometry Cystometry

Urethrogram Contrast enhanced ultrasound

Page 8: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Current mouse urinary phenotyping assays

can differentiate, but not diagnose

Female

Male

Strain 1 Strain 2 Strain 3 Strain 4

Inte

rva

l be

twe

en v

oid

s

Time

Bla

dd

er P

ress

ure

Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015

Control

Bladder outlet obstruction

*

**

Keil et al AJP Renal 2015

Inte

rva

l be

twe

en v

oid

sEnriched dietControl

Page 9: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

A mouse model of prostate inflammation

Ruetten et al In Press AJP-Renal

Collaboration between UW-Madison,

Columbia U. O’Brien Centers

Page 10: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Bacterial prostatitis does not always

increase urinary frequency

Ruetten et al AJP Renal In Press

Page 11: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Prostate inflammation causes different voiding

patterns in mice. Is this also true in humans?

Uninflamed

Prostate Inflamed

Bla

dde

r P

ressure

Ruetten and Bieberich Bell-Cohn, Thumbikat et al AJP Renal 2019

Vo

id F

reque

ncy

Vo

id F

reque

ncy

E. coli 1677

Lee, Bushman PLoS One 2015

E. coli CP1IL1 β overexpression

Ruetten et al In press

Obstructive

Irritative

Irritative

E. coli UTI89

Not easily binned

Page 12: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Maybe, we need to change our approach?

Many featuresSingle features

Nuclei + Sm. Muscle

+ Fibroblast + Inflammatory

Henry, Joseph, Strand et alKyle Wegner

Page 13: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Aggregate Void spot assay data successfully classifies mice based on disease process

• Hannah Ruetten, Gervaise Henry,

• Unpublished

• Collaboration between Vezina and Strand Teams

* Visit poster #47 for more information

Page 14: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

A phenomics approach for identifying LUTD

mechanisms and stratifying populations

Bacteria

(Strain, sequence)

Urinary

Phenome

Forward phenomics:

Pathogen phenotype

Reverse phenomics:

Phenotype to pathogen

Page 15: Segregating urinary phenome responses to microbes and ......Bjorling, Zeidel, Vezina, Hill et al AJP Renal 2015 Control Bladder outlet obstruction * * * Keil et al AJP Renal 2015 s

Doug Strand Lab, UT-Southwestern

Jill Macoska Lab, UT-Southwestern

Jonathan Barasch Lab, Columbia University

Chuck Bieberich, University of Maryland Baltimore

County

Dale Bjorling Lab, UW-Madison

Paul Marker Lab, UW-Madison

Will Ricke Lab, UW-Madison

Petra Popovics, UW-Madison

Matthew Grimes, UW-Madison

Collaborating Labs

Current

Thrishna Chathurvedula

Clara Cole

Olivia Fox

Thomas Peterson

Hannah Ruetten

Jaskiran Sandhu

Simran Sandhu

Brandon Scharpf

Anne Turco

Jonathan Zhu

Recent

Lisa Abler

Mark Cadena

Nicholas Girardi

Brett Mueller

Royal Oakes

Chelsea O’Driscoll

Kyle Wegner

Marlyse Wehber

Helen Zhang

obrien.urology.wisc.edu