>> HELLO, EVERYONE.
I WOULD LIKE TO WELCOME YOU TO
OUR SEMINAR TODAY, WHICH IS THE
OBESITY, NUTRITION AND PHYSICAL
ACTIVITY SEMINAR.
MY NAME IS CHARLOTTE PRATT.
I AM A DEPUTY BRANCH CHIEF IN
THE DIVISION OF CARDIOVASCULAR
SCIENCES AT THE NATIONAL HEART,
LUNG AND BLOOD INSTITUTE.
I WILL BE MODERATING THIS
SESSION, WHICH IS HOSTED BY THE
INSTITUTE'S OBESITY WORKING
GROUP.
PAST PRESENTATIONS HAVE
HIGHLIGHTED A VARIETY OF TOPICS
THAT HAVE BEEN ARCHIVED ON THIS
VIDEOCAST AND AVAILABLE TO THE
PUBLIC.
THESE INCLUDE NEW DIRECTIONS IN
COMMUNITY-BASED INTERVENTIONS TO
IMPROVE FOOD ACCESS AND REDUCE
OBESITY.
CARDIOVASCULAR DISEASE RISK, AND
INTERMITTENT FASTING AND THE
CIRCADIAN CLOCK, DOES WHEN YOU
EAT AFFECT YOUR HEALTH?
TODAY WE ARE SO FORTUNATE TO
HAVE DR. CASEY REBHOLZ PRESENT
ON DIETARY BIOMARKERS AND
CARDIOPROTECTIVE MECHANISMS OF
THE DASH DIET.
BEFORE I START, I WANT TO REVIEW
A FEW HOUSEKEEPING ITEMS.
THROUGHOUT THE PRESENTATION AND
DURING THE QUESTIONS AND ANSWER
DISCUSSIONS, PLEASE SEND YOUR
QUESTIONS TO THE EMAIL LISTED
BELOW ON SCREEN AT
ONPA@MAIL.NIH.GOV.
OR YOU MAY CLICK THE "LIVE
FEEDBACK" AT THE BOTTOM OF THE
VIDEOCAST SCREEN TO SUBMIT YOUR
QUESTION.
NOW LET ME INTRODUCE OUR
SPEAKER.
DR. CASEY REBHOLZ IS AN
ASSOCIATE PROFESSOR IN THE
DEPARTMENT OF EPIDEMIOLOGY AT
THE JOHNS HOPKINS BLOOMBERG
SCHOOL OF PUBLIC HEALTH.
SHE'S ALSO A FACULTY MEMBER AT
THE WELCH CENTER FOR PREVENTION,
EPIDEMIOLOGY AND CLINICAL
RESEARCH AND HAS A JOINT
APPOINTMENT IN THE DIVISION OF
NEPHROLOGY IN THE DEPARTMENT OF
MEDICINE AT JOHNS HOPKINS SCHOOL
OF MEDICINE.
HER RESEARCH FOCUSES ON
BIOMARKERS OF DIETARY INTAKE AND
CHRONIC DISEASE RISK.
DR. REBHOLZ IS CURRENTLY FROM
THE NHLBI AND IS CONDUCTING
RESEARCH USING ONE OF OUR MANY
COHORT STUDIES, THE
ATHEROSCLEROSIS RISK IN
COMMUNITIES, OR AS WE TYPICALLY
KNOW IT, ARIC.
HER RESEARCH FOCUSES ON
METABOLOMICS AND PROTEOMICS TO
IDENTIFY NEW AND OBJECTIVE
BIOMARKERS OF DIETARY INTAKE AND
DISEASE RISK.
PLEASE WELCOME DR. REB HOLDS. R.
>> THANK YOU SO MUCH, DR. PRATT,
FOR THE INTRODUCTION, AND THANK
YOU TO NHLBI FOR THE OPPORTUNITY
TO PRESENT TODAY.
SO HERE'S AN OUTLINE OF THE
TALK.
I'LL FIRST TALK ABOUT THE DASH
DIET AND HEALTH OUTCOMES
ASSOCIATED WITH THE DASH DIET,
AND THEN A LITTLE BIT ABOUT
DIETARY ASSESSMENT, AND THEN
INTO MY RESEARCH ON THE
METABOLOMICS OF THE DASH DIET IN
THE DASH TRIAL, IN THE DASH
SODIUM TRIAL, IN THE ARIC STUDY
AND IN THE CRIC STUDY AND THEN
FOLLOWING THAT, TALK ABOUT SOME
PROTEOMICS RESEARCH IN THE DASH
DIET IN THE ARIC STUDY AND THE
DASH SODIUM TRIALS, AND THEN
I'LL CONCLUDE.
SO WHAT IS THE DASH DIET?
IT'S A DIETARY PATTERN THAT'S
RICH IN FRUITS, VEGETABLES AND
LOW FAT DAIRY.
IT'S MODERATE IN A WIDE RANGE OF
PROTEIN SOURCES, AND LOW IN
SUGAR SWEETENED BEVERAGES,
SWEETS AND RED MEAT.
IT'S BEEN SHOWN IN CLINICAL
TRIALS TO REDUCE BLOOD PRESSURE
AND IN SUBSEQUENT EP DEEM YO
LICK STUDIES TO REDUCE THE RISK
OF HYPERTENSION, DIABETES,
STROKE, CARDIOVASCULAR DISEASE
AND MORTALITY.
AND IT'S BEEN WIDELY RECOMMENDED
AS HEALTHY DIETARY PATTERNS FOR
THE GENERATION POPULATION IN THE
U.S. DIETARY GUIDELINES FOR
AMERICANS AS WELL AS FOR THE
PREVENTION OF CARDIOVASCULAR
DISEASE BY THE AMERICAN HEART
ASSOCIATION.
AND HERE'S A FIGURE FROM A
RECENT SCIENTIFIC STATEMENT FROM
THE AMERICAN HEART ASSOCIATION
EVALUATING POPULAR DIETS.
AND THEY FOUND THAT THE
DASH-STYLE DIET WAS MOST ALIGNED
WITH THE LATEST DIETARY GUIDANCE
FROM THE AMERICAN HEART
ASSOCIATION.
SO WITH RESPECT TO HEALTH
OUTCOMES, HERE ARE THE FINDINGS
FROM THE ORIGINAL CLINICAL
TRIALS ON THE LEFT, THE DASH
TRIAL, AND ON THE RIGHT, THE
DASH SODIUM TRIAL.
SO YOU CAN SEE THAT AFTER THE
INITIAL TWO-WEEK PERIOD, THERE'S
A SUBSTANTIAL DROP IN BO BOTH
SYSTOLIC AND DIASTOLIC BLOOD
PRESSURE WHICH WAS SUSTAINED
OVER 8-WEEK PERIOD AND THIS
REDUCTION WAS MARKEDLY GREATER
FOR THE COMBINATION DIET
FOLLOWED BY THE FRUITS AND
VEGETABLES DIET COMPARED TO THE
CONTROL DIET.
SUBSEQUENT TO THE ORIGINAL
TRIAL, THE COMBINATION DIET HAS
BEEN REFERRED TO AS THE DASH
DIET.
ON THE RIGHT-HAND SIDE IN THE
DASH SODIUM TRIAL, WE HAVE
SIMILAR GRAPHS.
PARTICIPANTS WERE RANDOMIZED TO
THE DASH AND CONTROL DIET IN A
PARALLEL ARM DESIGN AND
SUBSEQUENTLY TO THREE SODIUM
LEVELS, HIGH, INTERMEDIATE AND
LOW, AND -- IN A CROSSOVER
DESIGN.
THOSE WHO EXPERIENCED THE
GREATEST REDUCTION IN BOTH
SYSTOLIC AND DIASTOLIC BLOOD
PRESSURE WERE FOLLOWING THE DASH
DIET DURING THE LOW SODIUM
INTERVENTION.
THEN BASED ON THE DESIGN OF THE
DASH DIET, SUBSEQUENT SCORES
HAVE BEEN CREATED TO ASSESS
ADHERENCE TO THE DASH DIET IN
FREE LIVING POPULATIONS.
ON THE LEFT IS A PRIMARILY FOOD
AND NUTRIENT FOCUSED SCORING
SYSTEM, AND ON THE RIGHT IS
PRIMARILY A NUTRIENT-FOCUSED
SYSTEM WITH MACRO AND MICRO.
SO ON THE LEFT, WE SEE THAT
HIGHER INTAKE OF SODIUM, RED AND
PROCESSED MEAT AND SUGAR
SWEETENED BEVERAGES RECEIVED
LOWER SCORES, AND HIGHER INTAKE
OF FRUITS, VEGETABLES, NUTS AND
LEGUMES, WHOLE GRAINS AND LOW
FAT DAIRY RECEIVE HIGHER SCORES.
ON THE RIGHT, WE SEE THAT LOWER
INTAKE OF SATURATED FAT, TOTAL
FAT, CHOLESTEROL AND SODIUM
RECEIVE HIGHER SCORES, AND
HIGHER INTAKE OF PROTEIN, FIBER,
MAGNESIUM, CALCIUM AND POTASSIUM
RECEIVED HIGHER SCORES ON THESE
TWO SCORING SYSTEMS.
SO HERE I'LL INTRODUCE THE ARIC
STUDY, WHICH I'LL MENTION AT
MANY POINTS THROUGHOUT THIS
PRESENTATION, WHICH DR. PRATT
ALREADY TALKED ABOUT.
BUT IT'S PRIMARILY AN NHLBI
COHORT, CERTAINLY AT ITS
INCEPTION IT HAS SUBSEQUENTLY
RECEIVED FUNDING FROM
DIFFERENT -- OTHER INSTITUTES.
IT WAS ESTABLISHED IN 1987 TO
1989, WAS THE VISIT 1, AT WHICH
POINT 15,792 MIDDLE AGED,
PREDOMINANTLY BUT NOT
EXCLUSIVELY WHITE AND BLACK MEN
AND WOMEN FROM FOUR U.S.
COMMUNITIES INDICATED WITH THE
RED STARS HERE ON THE MAP WERE
ENROLLED IN THE STUDY.
SUBSEQUENTLY THERE HAVE BEEN
MANY FOLLOW-UP VISITS THROUGHOUT
TIME.
THE FOOD FREQUENCY QUESTIONNAIRE
WAS ADMINISTERED AT BOTH VISIT Y
INTAKE.
AND I'LL MENTION SOME KIDNEY
FUNCTION SO I'VE NOTED HERE THAT
GFR IS A MEASURE OF KIDNEY
FUNCTION, WAS ESTIMATED AT MOST
OF THE FOLLOW-UP STUDY VISITS.
AND WE DEFINE INCIDENT CHRONIC
KIDNEY DISEASE AS A REDUCTION OF
GFR BELOW 60 AND AT LEAST A 25%
DECLINE RELATIVE TO BASELINE, AS
WELL AS ANY CKD RELATED
HOSPITALIZATION DEATHS OR ANY
INSTANCES OF END STAGE RENAL
DISEASE IDENTIFIED WITH THE
REGISTRY WHICH CAPTURES DIALYSIS
AND TRANSPLANT.
SO WE FOUND THAT WITH TERTILE
3 -- LOWER LEVELS OF ADHERENCE
WERE ASSOCIATED WITH HIGHER RISK
OF INCIDENT CHRONIC KIDNEY
DISEASE AND THIS ASSOCIATION
PERSISTED AFTER ADJUSTMENT FOR
MULTIPLE SOCIODEMOGRAPHIC,
HEALTH BEHAVIOR, KIDNEY
FUNCTION, BODY MASS, CLINICAL
CHARACTERISTICS AS WELL AS
HYPERTENSION LOWERING
MEDICATION.
AND THIS ASSOCIATION WAS
CONSISTENT WITH BOTH OF THE
SCORES THAT I PRESENTED, THE
FOOD-FOCUSED ONE AND THE KNEW
NUTRIENT FOCUSED ONE.
THIS WORK LED BY A FORMER PH.D.
STUDENT OF MINE, WE LOOKED AT
DASH DIET ADHERENCE AND
CARDIOVASCULAR DISEASE,
MORTALITY AND ALL CAUSE
MORTALITY.
WITH THE LOWEST QUINTILE AS THE
REFERENCE GROUP, NOW WE SEE THAT
HIGHER ADHERENCE IS ASSOCIATED
WITH A REDUCED RISK OF
CARDIOVASCULAR DISEASE MORTALITY
AND ALL CAUSE MORTALITY IN THE
OVERALL POPULATION.
AND SUGGESTION OF REDUCED RISK
OF CARDIOVASCULAR DISEASE AS
WELL.
AND THEN IN A DIFFERENT COHORT,
THIS IS THE CRIC STUDY OR
CHRONIC RENAL INSUFFICIENCY
COHORT STUDY FUNDED BY THE NIDDK
OF PEOPLE LIVING WITH KIDNEY
DISEASE, WIDE RANGE OF BASELINE
GFRs FROM 20 TO 70, IN THAT
POPULATION, AND WE FOUND THAT
HIGHER ADHERENCE TO THE DASH
DIET RELATIVE TO TERTILE
1 REPRESENTING THE LOWERED
ADHERENCE WAS ASSOCIATED WITH A
SIGNIFICANTLY REDUCED RISK OF
BOTH PROGRESSION OF CHRONIC
KIDNEY DISEASE AS WELL AS
ALL-CAUSE MORTALITY.
SO NOW A FEW THOUGHTS ON DIETARY
ASSESSMENT.
SO MEASURING DIET IS
CHALLENGING, AND IT'S IMPORTANT
THAT WE HAVE ACCURATE MEASURES
OF DIET FOR RESEARCH PURPOSES,
BOTH IN CLINICAL TRIALS AND
EPIDEMIOLOGIC STUDIES.
TYPICALLY WE USE SELF-REPORTED
DIETARY INTAKE, USING A FOOD
FREQUENCY QUESTIONNAIRE OR
24-HOUR DIETARY RECALL OR A FOOD
DIARY.
HOWEVER, THESE TOOLS ARE
SUBJECTIVE, AS WELL AS THE FOOD
COMPOSITION DATABASES THAT LINK
THE REPORTED INTAKE TO THE
NUTRIENTS WITHIN THOSE FOODS TO
ESTIMATE NUTRIENTS FOR A GIVEN
INDIVIDUAL, THEY ALSO HAVE THEIR
LIMITATIONS IN THAT THEY'RE
INCOMPLETE AND MAY BE
INACCURATE.
AND SO HEREIN LIES THE
ADVANTAGES OF DIET BIOMARKERS IN
THAT THEY'RE OBJECTIVE AND LESS
INFLUENCED BY BIAS AND ERROR,
SPECIFICALLY RECALL BIAS AND
SOCIAL DESIRABILITY BIAS.
BIOMARKERS ARE MORE PRECISE AND
LESS -- RANDOM ERRORS ARE LESS
OF A CONCERN.
HOWEVER, WITH DIETARY
BIOMARKERS, THE ONES THAT WE
HAVE TODAY THAT ARE CURRENTLY
USED REPRESENT PRIMARILY SINGLE
NUTRIENTS.
FOR EXAMPLE.
24-HOUR URINE EXCRETION OF
SODIUM TO ESTIMATE DIETARY
INTAKE OF SODIUM, POTASSIUM TO
ESTIMATE DIETARY INTAKE OF
POTASSIUM, AND UREA NITROGEN TO
ESTIMATE PROTEIN INTAKE.
HOWEVER, THE 24-HOUR URINE
COLLECTION IS BURDENSOME FOR
PARTICIPANTS, AND SO WHAT I'VE
DONE IS USED OMICS TECHNIQUES TO
DISCOVER NEW BIOMARKERS OF
DIETARY IB TAKE.
INTAKE.
THIS IS AN OVERVIEW OF
HIGH-THROUGHPUT TECHNOLOGIES
THAT ARE AVAILABLE TODAY IN
BIOLOGICAL RESEARCH RANGING FROM
THE MOST BROAD GENOMICS WHICH
MEASURES BIOLOGICAL POTENTIAL,
TRANSCRIPTOMICS WHICH
MEASURES -- AND THEN PROTEINS,
WHICH ARE ESSENTIAL FOR
BIOLOGICAL FUNCTION, AND THEN
FINALLY METABOLOMICS, WHICH ARE
ASSESSING ONE'S PHYSIOLOGIC
STATE, AND I'LL FOCUS TODAY ON
PROTEOMICS AND METABOLOMICS.
SO METABOLOMICS, SPECIFICALLY
GLOBAL OR UNTARGETED
METABOLOMICS PROFILING PROVIDES
AN UNBIASED AND COMPREHENSIVE
CHARACTERIZATION OF A COMPLEX
BIOSIS TEM.
BIOSYSTEM.
FURTHERMORE, ANOTHER ADVANTAGE
OF METABOLOMICS IS THE ABILITY
TO DETECT MANY SMALL COMPOUNDS,
AND THIS NUMBER IS INCREASING
EVERY DAY.
INTEGRATING THE METABALOME WITH
DIETARY DATA ALLOWS US TO LEARN
ABOUT FOOD INTAKE, AS WELL AS
FOOD METABOLISM.
AND IT COULD HELP US IDENTIFY
NOVEL BIOMARKERS FOR NUTRITION
RESEARCH, AS WELL AS
INTERVENTION TARGETS.
SO THIS PICTURE SHOWS YOU
INFLUENCES OF THE OVERALL
METABALOME, SO THERE ARE
COMPOUNDS THAT ARE IN THE BODY
ENDOGENOUSLY THAT ARE USED FOR
CELLULAR METABOLISM, AND THEN
THE FOOD METABALOME REPRESENTS A
WIDE RANGE OF FACTORS INCLUDING
INDIVIDUAL FOODS, OVERALL
DIETARY PATTERNS, FOOD ADDITIVES
AND CONTAMINANTS, DIETARY
SUPPLEMENTS, AND IN THE
METABALOME, WE'RE MEASURING
COMPOUNDS THAT ARE DERIVED FROM
THAT FOOD AFTER DIGESTION AND
METABOLISM.
THE METABALOME ALSO REPRESENTS
EXPOSURES TO DRUGS AND BROADER
ENVIRONMENTAL EXPOSURES SUCH AS
POLLUTANTS.
SO NOW ON TO METABOLOMICS OF THE
DASH DIET.
SO THIS, WE'LL BE STARTING WITH
THE DASH TRIAL, THE ORIGINAL
TRIAL.
HERE'S AN OVERVIEW OF THE DASH
TRIAL.
PARTICIPANTS FOLLOWED THE
CONTROL DIET FOR A THREE-WEEK
PERIOD AND THEN WERE RANDOMIZED
IN A PARALLEL ARM DESIGNED TO
EITHER THE CONTROL DIET, FRUITS
AND VEGETABLES DIET AND WHAT WAS
ORIGINALLY REFERRED TO AS THE
COMBINATION DIET BUT WE NOW CALL
THE DASH DIET FOR 8 WEEKS.
AND I OBTAINED THE SERUM
SPECIMENS FROM THE NHLBI'S
BIOLINCC REPOSITORY, A FABULOUS
RESOURCE.
THERE ARE 459 PARTICIPANTS
AVAILABLE WHO PARTICIPATED IN
THE ORIGINAL TRIAL AND A
SLIGHTLY SMALLER SUBSET WERE
AVAILABLE IN THE REPOSITORY FOR
THIS ANALYSIS.
THE METABOLOMICS PROFILING FOR
THIS STUDY WAS CONDUCTED BY
METABOL.
OH N, USING THEIR HD4 UNTARGETED
PLATFORM SO THIS USES FOUR
METHODS TO REVERSE PHASE ULTRA
PERFORMANCE LIQUID CHROME MA
TOGRAPHY TANDEM MASS
SPECTROMETRY WITH FOSTIVE ION
ELECTROSPRAY EYE ONIZATION,
REVERSE PHASE ULTRA PERFORMANCE
LIQUID CHROMATOGRAPHY WITH
NEGATIVE ION MODE ELECTROSPRAY
EYE ONIZATION HYDROPHILIC
INTERACTION YOU PRA PERFORMANCE
LICK YOU WID CHROMATOGRAPHY WITH
NEGATIVE ION MODE ELECTROSPRAY
EYE ONIZATION.
AND I PULLED 20 VIALS THAT WERE
MASKED TO THE LABORATORY, AND
WERE DUPLICATES OF THE ORIGINAL
VIALS, AND THE PERFORMANCE BY
THE LAB WAS EXCELLENT WITH A
MEDIAN CORRELATION OF .91, AND
72 METABOLITES HAVING A
CORRELATION ABOVE .8 WITHIN
THOSE 20.
A TOTAL OF 1238 METABOLITES WERE
DETECTED, AND FOR THIS, I
PRIMARILY FOCUSED ON THE KNOWN
METABOLITES EXCLUDING THE
UNKNOWNS, AND THOSE WERE 818
METABOLITES.
SO HERE ARE THE OVERALL RESULTS
FROM THIS STUDY.
97 SERUM KNOWN METABOLITES WERE
SIGNIFICANTLY DIFFERENT BETWEEN
THE DASH DIET AND THE CONTROL
DIET.
HERE WE HAVE A MODIFIED
MANHATTAN PLOT DISPLAYING THE
SIGNIFICANCE PLOTTED ALONG THE Y
AXIS AND CAT DWRIS OF THE
INDIVIDUAL METABOLITES ALONG THE
X AXIS.
AND THE HORIZONTAL LINE
REPRESENTS THE THRESHOLD FOR
STATISTICAL SIGNIFICANCE
ACCOUNTING FOR THE NUMBER OF
STATISTICAL COMPARISONS
CONDUCTED, THAT IS THE NUMBER OF
METABOLITES THAT WERE ANALYZED.
AND SO WE SEE A LARGE NUMBER OF
LIPIDS THAT WERE DIFFERENT AS
WELL AS AMINO ACIDS, XENOBIOTICS
WHICH INCLUDES FOOD AND
PLANT-DERIVED COMPOUNDS,
COFACTORS AND VITAMINS, A
CARBOHYDRATE AND A NUCLEOTIDE.
HERE ARE THE RESULTS FROM THE
PARTIALLY SQUARED DISKRIM NAT
ANALYSIS, AND WE SEE VERY CLEAR
SEPARATION IN THE PLOTS FOR THE
DASH AND THE CONTROL DIET, WITH
THE FIRST TWO COMPONENTS
EXPLAINING A LARGE PROPORTION OF
THE VARIABILITY, 29.4 FOR
COMPONENT 1 AND 12.6 FOR
COMPONENT 2.
AND FROM THIS ANALYSIS, WE WERE
ABLE TO IDENTIFY VARIABLE
IMPORTANCE AND PROJECTION
SCORES, WHICH CHARACTERIZES THE
MOST INFLUENTIAL METABOLITES FOR
DISCRIMINATING BETWEEN THE TWO
DIETS IN THIS STUDY, AND WE
FOUND SIX METABOLITES IDENTIFIED
HERE THAT WERE HIGHER ON THE
DASH DIET COMAIRED TO THE
CONTROL DIET AND THEN FOR
METABOLITES HIGHER ON THE
CONTROL DIET THAN THE DASH DIET.
IN A RANDOMLY SELECTED
TWO-THIRDS SAMPLE, THESE
METABOLITES WERE VERY IMPORTANT
IN PREDICTING THE DASH VERSUS
THE CONTROL DIET, AND SIMILARLY
FOR THE REMAINING ONE-THIRD
SAMPLE.
AND SO THESE TOP 10 METABOLITES
VERY MUCH REPRESENT THE KEY
FACTORS OF THE DASH DIET, SO
CHIRO-INOSITOL, STRUCTURALLY
RELATED TO FRUITS, BEANS,
GRAINS, NUTS AND SEEDS, AND N
METHYL PROLINE, CITRUS FRUIT AND
JUICE.
TRYPTOPHAN BETAINE, LEN TILS AND
LEGUMES.
GLUCOPYRANOSIDE, CAREER YELLS
AND CEREAL PRODUCTS AS WELL AS
TOTAL FRUIT INTAKE, B
CRYPTOXANTHIN, AND THE LAST FOUR
WERE -- THEOBROMINE,
7 METHYLXANTHINE,
3 METHYLXANTHINE, 7 METHYL
LURATE, CAFFE KA WHICH REPRESENT
CHOCOLATE HAVE BEEN ASSOCIATED
WITH LOWER DIET QAWLTD RELATED
TO COFFEE AND DESSERTS.
I'M GOING TO TRANSITION INTO THE
SECOND STUDY WITH IS URINE
METABOLOMICS OF THE DASH TRIAL
AND THE SECOND DASH SODIUM
TRIAL.
HERE'S THE OVERALL DESIGN IN
WHICH PARTICIPANTS FOLLOWED A
CONTROL DIET AND THEN WERE
RANDOMIZED TO EITHER THE DASH OR
THE CONTROL DIET AND THEN IN A
CROSSOVER DESIGN RECEIVED ONE OF
THREE DIFFERENT SODIUM
INTERVENTIONS.
FOR THIS ANALYSIS, I WAS
PARTICULARLY INTERESTED IN THE
DASH DIET COMPARED TO THE
CONTROL DIET.
BUT WE SELECTED DIFFERENT SODIUM
INTERVENTIONS, SO THE FIRST
COMPARISON IS THE DASH HIGH
SODIUM, VERSUS THE CONTROL HIGH
SODIUM.
SO THE ONLY DIFFERENCE HERE IS
THE DASH DIET, AND THEN THE DASH
LOW SODIUM VERSUS THE CONTROL
HIGH SODIUM TO SEE A MAXIMAL
CONTRAST BETWEEN THE DASH DIET,
WHICH IS CURRENTLY RECOMMENDED
AS THE LOW SODIUM DIET VERSUS A
TYPICAL AMERICAN DIET WHICH IS
HIGHER IN SODIUM.
AND WE USED URINE THAT'S
COLLECTED AT THE END OF EACH
INTERVENTION PERIOD AND OF THE
1425 METABOLITES IDENTIFIED
FOCUSED ON THE KNOWN METABOLITES
WHICH WERE 938.
AND HERE IN THIS PLOT ON THE
RIGHT-HAND SIDE, WE SEE THE
DISTRIBUTION OF THE KNOWN
METABOLITES THAT WERE DETEACTED
IDETECTEDIN THE PLATFORM BY PAT.
THEN FOR DASH HIGH SODIUM, THESE
ARE THE METABOLITES THAT WERE
SIGNIFICANTLY DIFFERENT FROM THE
CONTROL DIET, AND SIMILARLY FOR
THE DASH LOW SODIUM, THESE ARE
THE METABOLITES THAT WERE
SIGNIFICANTLY DIFFERENT BETWEEN
THE DASH LOW SODIUM DIET AND THE
CONTROL DIET.
SO THE DISTRIBUTION OF THE
CATEGORIES WAS SIMILAR ACROSS
THE COMPARISONS AND THE MOST
COMMON METABOLITE CATEGORY WAS
XENOBIOTICS, SO THESE FOOD
DERIVED COMPOUNDS.
AND HERE'S THE VOLCANO PLOT,
SHOWING THE SIGNIFICANT
METABOLITES THAT WERE DIFFERENT,
SO THE SIGNIFICANCE IS
IDENTIFIED AS THOUGHTS THAT
APPEAR -- DOTS THAT APPEAR ABOVE
THE RED DASHED LINE FOR THE DASH
HIGH SODIUM DIET ON PANEL A ON
THE LEFT AND FOR THE DASH LOW
SODIUM DIET IN PANEL B ON THE
RIGHT.
SO WE SEE -- AND THEN COLORS
REPRESENTING EACH OF THE
PATHWAYS.
SO WE SEE LOTS OF XENOBIOTICS
HERE ON BOTH SIDES WITH
POSITIVELY AND NEGATIVELY
ASSOCIATED WITH DASH DIET.
AND HERE ARE THE SCORES PLOTS
FROM THE PARTIALLY SQUARED
DISKRIM NAT ANALYSIS AGAIN SHOWS
VERY CLEAR DIFFERENTIATION
BETWEEN THE DASH DIET AND THE
CONTROL DIET FOR BOTH
COMPARISONS.
AND BASED ON THE VARIABLE
IMPORTANCE AND PROJECTION SCORES
WE'RE LISTING THE TOP 10
METABOLITES IN THE DASH SODIUM
STUDY FOR THE DASH HIGH SODIUM
CONTRAST AND THE DASH LOW SODIUM
CONTRAST, AND THE RESULTS HERE
ARE VERY SIMILAR ACROSS THE TWO
CONTRASTS.
WITH THREE OF THE METABOLITES
AND METHYL PROLINE -- BEING ALSO
AMONG THE TOP 10 IN THE DASH
TRIAL, WHICH I'VE PRESENTED
BEFORE.
AND HERE THE TOP 10 MOST
IMPORTANT METABOLITES WERE
EXCEEDINGLY INFLUENTIAL IN
PREDICTING THE INTERVENTION OF
THE DASH DIET.
IN BOTH TESTING AND VALIDATION
SAMPLES.
SO IN SUMMARY, IN THIS FEEDING
TRIAL OF ADULTS WITH HIGHER THAN
OPTIMAL BLOOD PRESSURE, WE
IDENTIFIED MANY METABOLITES
ASSOCIATED WITH THE DASH DIET
AFTER ADJUSTING FOR PARTICIPANT
CHARACTERISTICS AND ACCOUNTING
FOR MULTIPLE COMPARISONS.
AND SEVERAL OF THESE
ASSOCIATIONS WERE REPLICATED
COMPARED TO THE RESULTS I SHOWED
BEFORE IN THE DASH TRIAL.
THESE THREE METABOLITES AND
METHYL PROLINE, TRYPTOPHAN -- IN
THE TOP 10, AND THEN SIX TOTAL
METABOLITES, THREE ADDITIONAL
METABOLITES, WERE REPLICATED IN
THE TOP 15 FOR DASH SODIUM.
WITH THE EXCEPTION OF
GLUCOPYRANOSIDE AND B CRYPTO SAN
THIN, THEY WERE ENTIRELY
REPLICATED IN DASH SODIUM.
OF THE 97 METABOLITES THAT WERE
SIGNIFICANTLY DIFFERENT IN DASH,
17 AND 18 IN DASH HIGH SODIUM
AND DASH LOW SODIUM WERE ALSO
SIGNIFICANT AND SIMILAR IN
MAGNITUDE AS WELL AS IN
DIRECTION.
SO WITH THE FIRST STUDY BEING
CONDUCTED IN SERUM METABOLITES
AND THE SECOND IN URINE, THIS
PROVIDES SOME SUPPORT THAT THESE
MATRICES ARE POTENTIALLY
INTERCHANGEABLE FOR
CHARACTERIZING THE NUTRITION
METABALOME.
NOW BACK TO THE ARIC STUDY.
MOVING FROM A FEEDING TRIAL,
VERY WELL CONTROLLED ENVIRONMENT
TO A FREE LIVING POPULATION.
AGAIN, WE SEE THAT AT BASELINE,
VISIT 1, WE HAVE A NUMBER OF
PARTICIPANTS WHO COMPLETED THE
FOOD FREQUENCY QUESTIONNAIRE.
AND IN TERMS OF WHO WAS SELECTED
FOR METABOLOMICS PROFILING,
THERE WERE TWO SEPARATE, THE
FIRST ONE CONSISTED OF A RANDOM
SAMPLE OF ARIC PARTICIPANTS, AND
THE SECOND SUBGROUP CONSISTED OF
PARTICIPANTS WITH SEQUENCING
DATA.
AND THE OVERLAP IN TERMS OF THE
TWO METABOLOMICS PROFILING IN
THE TWO SUBGROUPS WAS A TOTAL OF
374 METABOLITES.
SO THIS ANALYSIS USES SUBGROUP P
2 FOR REPLICATION OF AGAIN THE
37 OVERLAPPING METABOLITES, AND
WE USE MULTIVARIABLE LINEAR
REGRESSION TO EXAMINE
ASSOCIATIONS BETWEEN DIETARY
PATTERNS AND INDIVIDUAL SERUM
METABOLITES ADJUSTING FOR
PARTICIPANT CHARACTERISTICS AND
WE ADJUSTED FOR MULTIPLE
COMPARISONS USING THE BON
FERRONI METHOD AND THEN META
ANALYZED SIGNIFICANT METABOLITES
ACROSS THE TWO SUBGROUPS USING
FIXED EFFECTS MODELS AND THEN
TESTED FOR PREDICTION WITH THE
ADDITION OF SIGNIFICANT
METABOLITES BEYOND PARTICIPANT
CHARACTERISTICS.
SO HERE WE SEE THAT 14
METABOLITES WERE SIGNIFICANTLY
ASSOCIATED WITH THE DASH DIET IN
THE ARIC STUDY IN SUBGROUP 1 AS
WELL AS SUBGROUP 2 AND THEY'RE
LISTED HERE: AMINO ACIDS,
CARBOHYDRATES, COFACTORS AND
VITAMINS, LIPIDS, AND A COUPLE
XENOBIOTICS.
FIVE OF THESE WERE ALSO
SIGNIFICANT IN THE DASH TRIAL,
SO N METHYL PRO NEEN, N ACETYL
ORNITHINE -- ADDITIONAL
METABOLITES WERE ALSO
SIGNIFICANT IN THE DASH SODIUM
TRIAL.
SO N METHYL -- IF YOU'LL
REMEMBER FROM THE DESIGN SLIDE,
THERE WERE MORE METABOLITES
IDENTIFIED IN SUBGROUP 2, SO
THERE WERE ADDITIONAL 20
METABOLITES THAT WERE ASSOCIATED
WITH THE DASH DIET, ONLY IN THIS
SUBGROUP.
AND MANY OF THOSE WERE
XENOBIOTICS AS WELL AS AMINO
ACIDS, CARBOHYDRATES, COFACTORS
AND VITAMINS AND LIPIDS.
SO OVERALL -- SO THIS ANALYSIS
WAS STUDYING THE OVERALL DASH
DIET, BUT THEN WITHIN THE ARIC
STUDY, WE HAVE THE BENEFIT OF
HAVING MORE DETAILED DIETARY
INFORMATION, SO WE LOOKED AT
FOOD GROUPS THAT CONTRIBUTE TO
THE DASH DIET SCORE, AND WE
INVESTIGATED OF THOSE
METABOLITES THAT ARE
SIGNIFICANTLY ASSOCIATED WITH
THE DASH DIET, WHICH FOOD
COMPONENTS OF THE OVERALL DASH
DIET SCORE ARE THEY ASSOCIATED
WITH?
SO YOU SEE N METHYLPROLINE ALSO
ASSOCIATED WITH FRUITS.
AND YOU CAN GO DOWN THE LIST.
BUT IT ALIGNS WELL WITH THE --
THE METABOLITES ASSOCIATED WITH
THE DASH DIET ALIGN WELL WITH
THE KEY COMPONENTS OF THE DASH
DIET SCORE AND IT HELPS US
INTERPRET THESE FINDINGS.
IN TERMS OF WHICH SPECIFIC FOOD
GROUPS THEY REPRESENT.
AND THEN WE SEE THAT THE
ADDITION OF THE METABOLITES
SIGNIFICANTLY IMPROVE PREDICTION
OF HIGH ADHERENCE TO THE DASH
DIET BEYOND THE PARTICIPANT
CHARACTERISTICS IN BOTH SUBGROUP
1 AND SUBGROUP 2.
SO IN SUMMARY, WE IDENTIFIED 14
METABOLITES ASSOCIATED WITH DASH
DIET IN TWO DIFFERENT SUBGROUPS
WITHIN THE ARIC STUDY, AND
ANOTHER 20 METABOLITES THAT
SERVE AS CANDIDATE BIOMARKERS
THAT WERE ONLY AVAILABLE IN THAT
SECOND SUBGROUP.
AND THEN OF THESE FINDINGS,
THERE WAS REPLICATION ACROSS
BOTH THE TWO SUBSAMPLES WITHIN
THE ARIC STUDY AS WELL AS THE
DASH TRIAL AND THE DASH SODIUM
TRIAL, SO N METHYL PROLINE, 3
3-4 HYDROXYPHENYL ACETATE, N
ACETYL LORN THEEN, PEER
DOCKSITE, GAMMA -- THESE
METABOLITES WERE ASSOCIATED WITH
FOODS INCLUDING FRUIT,
VEGETABLES, WHOLE GRAINS, NUTS
AND LEGUMES, DAIRY, RED MEAT,
SUGAR SWEETENED BERCHLS AND
SODIUM, SO ALL THE KEY
COMPONENTS.
AND THEN THESE METABOLITES
SIGNIFICANTLY IMPROVED ADHERENCE
TO THE DASH DIET.
SO NOW COMING BACK AGAIN TO THE
CRIC STUDY, THIS COHORT STUDY OF
PARTICIPANTS LIVING WITH KIDNEY
DISEASE.
SO AS BASELINE, WHICH WAS IN
2003 TO 2008, 3,930 PARTICIPANTS
WERE ENROLLED, WIDE RANGE OF
AGES, 21 TO 74 AND A WIDE RANGE
OF KIDNEY FUNCTIONS, 20 TO 70.
ONLY A SUBSET OF THESE HAD A
DIET HISTORY QUESTIONNAIRE THAT
WAS ADMINISTERED AND
METABOLOMICS DATA THAT WAS
AVAILABLE CONDUCTED IN PLASMA.
IN THIS STUDY, THE METABALOME
PROFILING WAS CONDUCTED AT THE
BROAD INSTITUTE IN BOSTON, AND
YOU CAN SEE THAT THE PLATFORM IS
QUITE DIFFERENT FROM THE DATA
WHICH WAS GENERATED FROM THE
BROAD INSTITUTE.
WE TRY TO CLASSIFY THE
METABOLITES IN A SIMILAR FASHION
AS IN THE PREVIOUS ANALYSIS.
AND WE SEE THAT THIS PLATFORM
SPECIFICALLY HAS A LARGE
REPRESENTATION OF LIPIDS AND
LIPID SUBCLASSES.
SO IN TERMS OF THE ANALYSIS, WE
RAN MULTIVARIABLE LINEAR
REGRESSION MODELS TO EXAMINE
ASSOCIATIONS BETWEEN DASH DIET
SCORES AND INDIVIDUAL PLASMA
METABOLITES, ADJUSTING FOR
SOCIODEM GRARVEG
CHARACTERISTICS, HEALTH
BEHAVIORS AND CLINICAL FACTORS
ADJUSTING FOR MULTIPLE FACTORS
USING THE BONFERRONI METHODS.
THEN WE ALSO USED PRINCIPAL
COMPONENTS ANALYSIS TO LOOK AT
OF THE METABOLITES WHICH WERE
SIGNIFICANTLY ASSOCIATED WITH
THE DASH DIET, HOW WERE THESE
FORM IN PRINCIPLE COMPONENTS,
HOW DO THEY FALL TOGETHER IN
PRINCIPAL COMPONENTS AND WHICH
FOOD COMPONENTS ARE THEY RELATED
TO OF THE COMPONENTS REPRESENTED
WITHIN THE DASH DIET?
AND WE RETAINED FIVE COMPONENTS
FOR THE DASH DIET WHICH TOGETHER
EXPLAINED 64% OF THE VARIANTS.
VARIANCE.
HERE IN THE GRAPH ON THE
RIGHT-HAND SIDE, WE CAN SEE A
SIMILAR PICTURE AS WHAT WE SAW
BEFORE, BUT AGAIN, MANY MORE
SUBCLASSES OF LIPIDS IN THE
IDENTIFIED METABOLITES BY THE
PLATFORM, AND THEN ON THE
LEFT-HAND SIDE, THOSE THAT WERE
SIGNIFICANTLY ASSOCIATED WITH
THE DASH DIET.
AND THERE WERE A TOTAL OF
77 THAT WERE ASSOCIATED WITH THE
DASH DIET, AND THE MAJOR
CATEGORIES HERE ARE
TRIACYLGLYCEROLS IN THE BRIGHT
GREEN COLOR AND THEN PCs,
PEs AND PSs, SO THAT IS IN
THE ROYAL BLUE, WHICH WERE 13%
OF THE SAMPLE AND THEN THE
TRIACYLGLYCEROLS WERE AT 39% OF
THE SIGNIFICANT METABOLITES
ASSOCIATED WITH THE DASH DIET.
AND HERE IS A VOLCANO PLOT OF
THE BETA COEFFICIENTS FOR THE
ASSOCIATION OF THE INDIVIDUAL
METABOLITES WITH THE DASH DIET
ALONG THE X AXIS AND THE
NEGATIVE LOG TIME KEY VALUES
PLOTTED ALONG THE Y AXIS WITH
THE VERTICAL RED DASHED LINE
REPRESENTING THE NULL VALUE OF
ZERO, AND THE HORIZONTAL RED
DASHED LINE REPRESENTING THE
BONFERRONI ADJUSTED THRESHOLD
FOR STATISTICAL SIGNIFICANCE.
SO TO THE LEFT -- UPPER
LEFT-HAND SIDE, WE SEE
METABOLITES THAT WERE INVERSELY
ASSOCIATED WITH THE DASH DIET,
AND MANY OF THESE ARE GREEN SO
TRY ACLGLIGLYCEROLS AND
DIACYLGLYCEROLS IN THE BLUE.
AND THEN THOSE THAT WERE
POSITIVELY ASSOCIATED WE SEE
SOME AMINO ACIDS IN RED AND
CHOLESTEROL ESTERS IN YELLOW.
WE ALSO LOOKED AT -- GIVEN THE
PREDOMINENCE OF LIPIDS IN THIS
PLATFORM, WE DECIDED TO LOOK AT
ASSOCIATIONS WITH ACYL CHAIN
LENGTH AS WELL AS DOUBLE BONDS,
AND WE FOUND THAT AS YOU GO
ALONG THIS X AXIS ON THE
LEFT-HAND SIDE PANEL, YOU HAVE
LONGER CHAIN LENGTH, AND MORE
STRONGLY NEGATIVE BETA
COEFFICIENTS, WHICH IS
INTERPRETED AS LIPIDS WITH
LONGER CHAIN LENGTH WERE
INVERSELY ASSOCIATED WITH THE
DASH DIET.
AND THEN YOU SEE SOME POSITIVE
ASSOCIATIONS FOR SOME OF THESE
VERY LONG CHAINS, SO WE
INTERPRETED THIS AS LONGER TRY
ASELLGLYCEROLS WERE POSITIVELY
ASSOCIATED WITH THE DASH DIET.
THEN IF HE WITH LOOK AT THE
PANEL ON THE RIGHT-HAND SIDE OF
DOUBLE BONDS, WE SEE IN GREEN
THE TRY ACYLGLYCEROLS THAT HAD
SEVEN OR MORE DOUBLE BONDS WERE
POSITIVELY ASSOCIATED WITH THE
DASH DIET.
THESE ARE THE RESULTS OF THE
PRINCIPAL COMPONENTS ANALYSIS.
SO WE'RE PRESENTING THE RESULTS
FROM FOUR PRINCIPAL COMPONENTS,
SO THE FIRST PRINCIPAL COMPONENT
WAS LOW IN DAGs AND TAGs
WITH A FEW DOUBLE BONDS.
THIS WAS POSITIVELY ASSOCIATED
WITH TOTAL VEGETABLES AND NUTS
AND LEGUMES.
THE SECOND PRINCIPAL COMPONENT
WAS LOW AND THIS WAS POSITIVELY
ASSOCIATED WITH RED AND
PROCESSED MEAT, INDICATING LOWER
INTAKE JUST BECAUSE OF HOW THE
SCORING SYSTEM WORKS AND THEN
THE THIRD PRINCIPAL COMPONENT
CONSISTED -- WAS LOW IN
COFACTORS AND VITAMINS, AND THIS
WAS INVERSELY ASSOCIATED WITH
SUGAR SWEETENED BEVERAGES,
REPRESENTING HIGHER INTAKE.
SUGAR SWEETENED BEVERAGES AND
THE PRINCIPAL COMPONENT 4 WAS
LOW -- THAT WAS INVERSELY
ASSOCIATED WITH TOTAL
VEGETABLES.
SO IN SUMMARY WE FOUND IN THE
CRIC STUDY, 77 METABOLITES
ASSOCIATED WITH THE DASH DIET
AND LIPIDS WITH LONGER CHAIN
LENGTH AND MORE DOUBLE BONDS SO
THE MORE UNSATURATED THEY WERE,
THE MORE POSITIVELY ASSOCIATED
WITH THE DASH DIET.
SATURATED FATS, TAGs AND
DAGs WERE INVERSELY ASSOCIATED
WITH HEALTHY FOODS, VEGETABLES
BE NUTS AND LEGUMES, AND
UNSATURATED TAGs, THOSE WITH 7 E
POSITIVELY ASSOCIATED WITH
VEGETABLE INTAKE.
SO COME BACK TO THE SUMMARY SIDE
OF HIGH-THROUGHPUT TECHNOLOGIES
WE'VE TALKED A LOT ABOUT THE
METABOLOMICS OF THE DASH DIET
AND I'M GOING TO TRANSITION NOW
INTO THE PROTEOMICS OF THE DASH
DIE E SO STARTING WITH
PROTEOMICS OF THE DASH DIET IN
THE ARIC STUDY.
THIS WORK WAS LED BY A CURRENT
PH.D. STUDENT OF MINE, SHUTONG
DU, AND IT WAS CONDUCTED IN THE
ARIC STUDY USING A SOMALOGIC
ASSAY THAT USES CHEMICALLY
MODIFIED NUCLEOTIDES, APTAMERS,
AS PROTEIN BINDING REAGENTS.
WE HAVE FOUR THOWN 955 PROTEINS
AVAILABLE FOR ANALYSIS.
AGAIN IN THESE MASS DUPLICATES,
THE CV WAS VERY LOW, AND THE
CORRELATION OR RELIABILITY
COEFFICIENT WAS HIGH.
WHICH MEANS THE ASSAY WORKED
WELL, HIGH PERFORMING ASSAY,
HIGH QUALITY ASSAY.
AND HERE AGAIN IS THE VOLCANO
PLOT WITH THE BETA COEFFICIENT
FOR THE ASSOCIATION OF PROTEINS
WITH THE DASH DIET ALONG THE X
AXIS AND THEN LOG 10P VALUES
REPRESENTING STATISTICAL CYST
CANS ALONG THE Y ACCESS.
WE SEE A TOTAL OF 254 PROTEINS
THAT WERE SIGNIFICANTLY
ASSOCIATED WITH THE DASH DIET AT
THE BONFERRONI THRESHOLD, P
VALUES BELOW 1 TIMES 10 TO THE
NEGATIVE 5TH AND THESE MODELS
WERE ADJUSTED FOR A NUMBER OF
SOCIODEMOGRAPHIC, SOCIOECONOMIC
HEALTH BEHAVIORS, BODY MASS
INDEX AND CLINICAL
CHARACTERISTICS.
SO WE SEE EIGHT OF THESE
METABOLITES THAT IMPROVED THE
PREDICTION OF HIGH ADHERENCE TO
THE DASH DIET BEYOND PARTICIPANT
CHARACTERISTICS.
AND IN PATHWAY ANALYSIS, THE
COMPLEMENT AND COAGULATION
CASCADE AND CELL ADHESION
MOLECULES WERE OVERREPRESENTED.
SO WHAT DO ALL THESE FINDINGS
MEAN?
WELL, I'LL HIGHLIGHT A FEW KEY
PROTEINS WE FOUND.
SO ONE IS EPIDERMAL GROWTH
FACTOR RECEPTOR, SO THIS IS
INVOLVED IN INFLAMMATION
ASSOCIATED WITH ANGIOGENESIS,
ATHEROSCLEROSIS AND DIABETES.
INSULIN-LIKE GROWTH FACTOR
BINDING PROTEIN IS ASSOCIATED
WITH LOW LEVELS OF -- IMPAIRED
GLUE COST TOLERANCE,
CARDIOVASCULAR DISEASE AND
HYPERTENSION, HIGHER LEVELS
PROMOTE NITRIC ACID PRODUCTION.
CONTACTIN-1 IS A CELL ADHESION
PROTEIN AND HAS BEEN INVERSELY
ASSOCIATED WITH MYOCARDIAL
INFARCTION, HEART FAILURE AND
DEATH.
NEURONAL GROWTH FACTOR 1 WAS
ASSOCIATED IN OTHER STUDIES WITH
BODY WEIGHT REGULATION, VIA THE
HYPOTHALAMUS, WHICH CONTROLS
FOOD CONSUMPTION, ESPECIALLY
FOODS THAT ARE HIGHER IN CARBS
AND FIBER AND LOWER IN FAT.
AND ALSO LOW LEVELS OF THIS
PROTEIN WERE ASSOCIATED WITH
ABNORMAL FAT DISTRIBUTION.
NEUROEXOPHILIN-1 HAS BEEN SHOWN
TO REGULATE MANY CARDIOVASCULAR
PARAMETERS, INCLUDING BLOOD
PRESSURE, LDL CHOLESTEROL, HDL
CHOLESTEROL AND ASSOCIATED WITH
DIABETES.
ENDO NUCLEOSIDE TRIPHOSPHATE
INVOLVED IN BODY WEIGHT REG EU
LAITION, ENERGY METABOLISM AND
INSULIN SENSITIVITY.
NEUROPEPTIDE W IS A WELL-KNOWN
HORMONE INVOLVED IN REGULATING
FOOD CONSUMPTION AND ENERGY
HOMEOSTASIS IN PERIODS OF
STRESS.
SO SOME REALLY INTERESTING
FINDINGS THAT ALIGN WELL WITH
THE BASIC SCIENCE RESEARCH AND
SOME EPIDEMIOLOGIC RESEARCH
LOOKING AT CLINICAL OUTCOMES
ASSOCIATED WITH THESE PROTEINS.
SO THE PATHWAYS THAT WE
IDENTIFIED THAT WERE
OVERREPRESENTED AMONG THE
SIGNIFICANT PROTEINS WERE THE
COMPLEMENT AND COAGULATION
PROCESSES THAT ARE INVOLVED IN
INNATE IMMUNITY, TISSUE INJURY
AND INFLAMMATORY RESPONSE, AND
THEN CELL ADHESION MOLECULES
INVOLVED IN HEMOYOE STASIS,
IMMUNE RESPONSE, INFLAMMATION
AND ATHEROSCLEROSIS.
SO THE LAST PROJECT I'LL MENTION
TODAY IS ON THE PROTEOMICS OF
THE DASH DIET IN THE TWO FEEDING
TRIALS, THE ORIGINAL DASH TRIAL
AND THEN THE FOLLOWING DASH
SODIUM TRIAL.
SO HERE WE COLLECTED SERUM
SPECIMENS AND THESE WERE
OBTAINED AGAIN FROM THE NHLBI
BIOLINCC REPOSITORY.
FROM THE END OF THE DASH DIET
WITHIN THE DASH TRIAL AND THE
DASH SODIUM TRIALS.
THIS VERSION OF THE SOMALOGIC
PLATFORM IDENTIFIED 7,241
PROTEINS, AND THEN PROTEINS THAT
WERE STATISTICALLY SIGNIFICANT
IN THE META-ANALYSIS ACROSS THE
TWO FEEDING TRIALS, SO DASH AND
DASH SODIUM, WERE THEN EVALUATED
IN THE ARIC STUDY.
SO HERE'S THE VOLCANO PLOT, AND
YOU'VE SEEN THIS DISPLAYED
BEFORE.
WE FOUND 71 PROTEINS THAT WERE
DIFFERENT BETWEEN THE DASH AND
CONTROL DIET, FROM THE
META-ANALYSIS ACROSS THE TWO
TRIALS.
AND THESE MODELS WERE ADJUSTED
FOR AGE, SEX, RACE, ETHNICITY,
AND BODY MASS INDEX, AND WE USED
AN FDR CUT POINT OF .05 TO
ASSESS STATISTICAL SIGNIFICANCE
FOR THIS PROJECT.
AND THEN OF THE 71, 50 WERE
AVAILABLE AND DETECTED IN THE
ARIC STUDY.
OF THOSE 50, 19 WERE ALSO
SIGNIFICANTLY ASSOCIATED WITH
THE DASH DIET IN THE ARIC STUDY,
AND WE CALL THOSE VALIDATED.
SO ON THE LEFT-HAND SIDE, YOU
SEE THESE 19 METABOLITES AND THE
BETA COEFFICIENTS -- SORRY, NOT
METABOLITES, PROTEINS, FOR
ASSOCIATION OF THESE PROTEINS
WITH THE DASH DIET IN THE
META-ANALYSIS OF THE DASH TRIAL,
IN THE DASH SODIUM TRIAL.
IN THE RIGHT-HAND SIDE PANEL,
YOU SEE RESULTS FROM THE ARIC
STUDY.
SO THESE METABOLITES WERE NOT
ONLY STATISTICALLY SIGNIFICANT
IN ALL THREE TRIALS, ALL THREE
STUDY POPULATIONS, BUT ALSO WERE
VERY CONSISTENT IN TERMS OF
THEIR DIRECTION OF ASSOCIATION.
SO WE FOUND 19 PRO T PROTEINS TT
COLLECTIVELY IMPROVED THE
PREDICTION OF THE DASH DIET IN
THE FEEDING STUDIES, AND THE
DASH DIET SCORE ASSOCIATED WITH
ADHERENCE TO THE DASH DIET AND
THE ARIC STUDY BEYOND
PARTICIPANT CHARACTERISTICS.
SO TO INTERPRET SOME OF THESE
PROTEIN NAMES, RBP4 IS A CARRIER
PROTEIN FOR VITAMIN A, AND IT'S
DERIVED FROM FRUITS AND
VEGETABLES.
FOLR2 BINDS TO FOLATE AND
DELIVERS 5 METHYL
TETRAHYDROFOLATE TO THE CELL.
AFM IS A VITAMIN E BINDING
PROTEIN, POSITIVELY ASSOCIATED
WITH ADIPOSITY, METABOLIC
SYNDROME, FATTY LIVER AND
DIABETES.
I MEMBERS OF THE JURIED EGFR,
INVOLVED IN CELL ADHESION FOR
STABILITY OF PLAQUES AND
INVERSELY ASSOCIATED WITH
CARDIOVASCULAR DISEASE.
THERE WERE SEVERAL PROTEINS THAT
HAVE A ROLE IN THE INFLAMMATORY
PROCESS.
AND THEN SEVERAL PROTEINS THAT
ARE INVOLVED IN KOL JENS
DEPOSITION, AND LOWERING
ARTERIAL STIFFNESS.
AND THEN CD163, WHICH IS A
TRANSMEMBRANE SCAVENGER RECEPTOR
PROTEIN IN MACROPHAGES AND
MONOCYTES INVOLVED IN
INFLAMMATION AND IS ASSOCIATED
WITH BOTH DIABETES AND OBESITY.
IN OTHER RESEARCH.
SO I'LL MOVE INTO THE CONCLUSION
NOW, SO IN SUMMARY, FROM THIS
LAST PROJECT, WE IDENTIFIED 19
PROTEINS THAT WERE ROBUSTLY
ASSOCIATED WITH THE DASH DIET IN
THREE STUDIES.
IN THE DASH TRIAL, DASH SODIUM
TRIAL AND ARIC STUDY.
THESE PROTEINS REFLECTED DIETARY
INTAKE, AS WELL AS LOWER
INFLAMMATION AND COLLAGEN
DEPOSITION.
OTHER PATHWAYS AS WELL, BUT THIS
IS KIND OF -- THERE WAS MANY
PROTEINS INVOLVED IN THESE
SPECIFIC PATHWAYS.
AND SO OUR FINDINGS SUGGEST
POTENTIAL PATHWAYS THAT ARE
IMPACTED BY CONSUMING THE DASH
DIET.
SO OVERALL FROM TODAY'S TALK,
THE DASH DIET IS A KNOWN HEALTHY
DIETARY PATTERN THAT HAS BEEN
SHOWN TO REPEATEDLY REDUCE BLOOD
PRESSURE LEVELS, AS WELL AS
REDUCE THE RISK OF INCIDENT
CHRONIC KIDNEY DISEASE,
CARDIOVASCULAR DISEASE MORTALITY
AND ALL-CAUSE MORTALITY IN
HEALTHY INDIVIDUALS, AS WELL AS
KIDNEY DISEASE PROGRESSION I AND
MORTALITY IN KIDNEY PATIENTS, SO
THESE ARE THE RESULTS FROM THE
ARIC STUDY AND FROM THE CRIC
STUDY.
FOLLOWING THE FINDINGS FROM THE
ORIGINAL CLINICAL TRIALS.
NUTRITIONAL METABOLOMICS IS A
USEFUL APPROACH FOR DISCOVERY OF
DIETARY BIOMARKERS.
AND METABOLOMICS MARKERS OF THE
DASH DIET FROM FEEDING TRIALS,
AS WELL AS COHORT STUDIES, COULD
BE USED IN FUTURE STUDIES.
I ALSO TALKED ABOUT PROTEOMICS,
SO I THINK PROTEOMICS OFFERS
COMPLEMENTARY INFORMATION BEYOND
METABOLOMICS THAT POINTS TO KEY
PATHWAYS THAT UNDERLINE -- THAT
UNDERLIE THE DIET-DISEASE
RELATIONSHIP AND PATHWAYS THAT
ARE RELEVANT FOR FOOD DIGESTION,
ABSORPTION AND METABOLISM.
I DIDN'T LINK THE PROTEOMICS TO
DISEASE OUTCOMES TODAY, BUT
THOSE FINDINGS ARE COMING, AND I
THINK WE CAN INFER THIS BASED ON
WHAT WE KNOW ABOUT THE PROTEINS
FROM OTHER RESEARCH.
SO WHAT'S NEXT?
THIS IS REALLY A DISCOVERY-BASED
APPROACH TO IDENTIFYING NEW
MARKERS OF DIETARY INTAKE, SO
THERE'S A LOT MORE THAT NEEDS TO
BE DONE.
SO SOME OF THE FUTURE RESEARCH
THAT I HAVE PLANNED IS TO LOOK
AT THESE DASH DIET RELATED
METABOLITES AND PROTEINS AND
LINK THEM TO CLINICAL
PARAMETERS, SO MY FIRST INTEREST
IS BLOOD PRESSURE AND
HYPERTENSION, BUT ALSO OTHER
OUTCOMES.
SECOND, TO FOLLOW UP ON THESE
CANDIDATE MARKERS, I AM
DEVELOPING QUANTITATIVE ASSAYS,
AND THEN WE'LL BE TESTING THEM
USING SPECIMENS.
AND THEN THE DASH DIET CLEARLY
IS VERY IMPORTANT, IS WIDELY
PROMOTED, AND IS RELEVANT
SPECIFICALLY FOR THE U.S.
POPULATION.
BUT THERE'S OTHER DIETARY
PATTERNS OF INTEREST, AND
THERE'S OTHER DIETARY FACTORS OF
INTEREST.
SO THERE'S MORE WORK TO DO TO
BUILD THIS EVIDENCE ON
BIOMARKERS OF DIET.
I'D LIKE TO MAKE A FEW
ACKNOWLEDGMENTS, PRIMARILY
FUNDING, FROM THE NHLBI.
SO I HAVE THE BENEFIT OF HAVING
BEEN FUNDED BY THE NHLBI A FEW
TIMES IN THIS INITIAL R21 WAS TO
ENCOURAGE USE OF THE BIOLINCC
REPOSITORY, WHICH I MENTIONED A
COUPLE TIMES TODAY, AND THEN
THIS R56 HIGH IMPACT ONE-YEAR
AWARD FOLLOWED BY MY FIRST R01,
SO I'M VERY GRATEFUL FOR
FUNDING.
I ALSO WOULD LIKE TO ACKNOWLEDGE
ALL THE PARTICIPANTS WHO WERE
INVOLVED IN THESE STUDIES, AND
THE MANY PEOPLE WHO CONTRIBUTED
TO THIS RESEARCH AT MANY STAGES.
I DON'T HAVE THEM ALL LISTED
BECAUSE I RISK LEAVING SOMEONE
OFF, BUT THERE ARE MANY PEOPLE.
SO THAT'S ALL I HAVE FOR TODAY.
I WELCOME ANY QUESTIONS THAT
YOU HAVE.
>> THANK YOU SO MUCH,
DR. REBHOLZ.
THIS WAS FANTASTIC!
YOUR PRESENTATION REALLY CALLED
ATTENTION TO THE NEED FOR
OBJECTIVE MEASURES OF DIETARY
INTAKE, AND YOU REALLY TOUCHED
BASE ON SEVERAL COMPONENTS, THE
DASH DIET, AS WELL AS THE ARIC
STUDY AND THE CHRONIC KID NIGH
DISEASE KIDNEYDISEASE STUDY, SOO
MUCH.
JUST A REMINDER, PLEASE PUT YOUR
QUESTIONS IN THE CHAT BOX OR
SEND TO THE ONPA LINK ON YOUR
SCREEN, IT'S THE
ONPA@MAIL.NIH.GOV.
AND WE WILL BE COLLECTING
QUESTIONS SO THAT DR. REBHOLZ
CAN PROVIDE ANSWERS TO THOSE
QUESTIONS.
SO TO START US OFF, I HAVE ONE
QUESTION.
I NOTICED STRONG VARIABILITY IN
THE PLOTS THAT YOU PROVIDED, AND
I WAS WONDERING WHETHER YOU HAVE
SOME DIRECTIONS IN TERMS OF
PERHAPS MAYBE WE NEED TO LOOK AT
PRECISION NUTRITION BECAUSE OF
THIS KIND OF VARIABILITY IN THE
PLOTS, EVEN WITH THE
METABOLITES.
SO I WONDERED IF YOU HAVE ANY
RESPONSE TO THAT.
>> YEAH, I THINK, YOU KNOW,
OVERALL, WE WERE ABLE TO FIND
VERY CONSISTENT ASSOCIATIONS
ACROSS THE FEEDING TRIALS, AS
WELL AS TRANSLATING TO COHORT
STUDIES, WHICH IS A VERY
DIFFERENT DESIGN, VERY DIFFERENT
SETTING, PEOPLE WEREN'T
RECOMMENDED TO FOLLOW THE DASH
DIET, SO THAT'S MY TAKE-HOME,
THAT THE RESULTS ARE QUITE
CONSISTENT.
ACROSS THE STUDIES.
AND I THINK IT'S USEFUL, AGAIN,
TO FIND BIOMARKERS THAT, ON
AVERAGE, ASSESS -- ARE USED FOR
INDICATING ADHERENCE TO THE DASH
DIET AND OTHER DIETARY FACTORS.
BUT YOUR POINT IS A GOOD ONE,
YOU KNOW, IT IMPORTANT TO
UNDERSTAND, YOU KNOW, ARE THESE
UNIVERSAL OR DO THEY DIFFER FOR
CERTAIN SUBGROUPS OF THE
POPULATION.
AND WE HAVE STARTED DOING THAT,
LOOKING AT SIMILAR -- IF THE
ASSOCIATIONS ARE SIMILAR BY --
BETWEEN MEN AND WOMEN, BETWEEN
RACIAL/ETHNIC GROUPS AS WELL,
AND LARGELY -- I DIDN'T PRESENT
THIS, BUT LARGELY THE RESULTS
ARE VERY CONSISTENT.
BUT THAT ISN'T TO SAY, YOU KNOW,
THERE IS VARIABILITY AND IT
NEEDS TO BE UNDERSTOOD, AND SO I
THINK THAT IS A VERY IMPORTANT
NEXT STEP AND APPROACH TO TRYING
TO FIND SOME MEANING BEHIND
THESE DATA.
>> THANK YOU SO MUCH.
SEVERAL QUESTIONS HAVE COME UP.
THERE'S A QUESTION SOMEONE SENT
WANTS TO KNOW, YOU MENTIONED
THEOBROMINE WAS IDENTIFIED IN
THE CLINICAL TRIAL, AND THIS
METABOLITE IS ALSO ASSOCIATED
WITH CHOCOLATE AND LOWER DIE DIY
QUALITY.
SO DO YOU KNOW IF THIS
METABOLITE HAS ASSOCIATION WITH
CARDIOMETABOLIC HEALTH?
>> YEAH, THAT'S A GREAT
QUESTION.
SO FOR TODAY, I WAS MOSTLY
FOCUSING ON, YOU KNOW, WORK
TRYING TO UNDERSTAND BIOMARKERS
OF DIET WHICH I THINK IS AN
IMPORTANT GOAL IN AND OF ITSELF,
BUT OF COURSE, YOU KNOW, A MAJOR
BENEFIT OF THESE TECHNIQUES,
METABOLOMICS AND PROTEOMICS, IS
THAT THEY CAN HELP POINT US IN
THE RIGHT DIRECTION IN TERMS OF
UNDERSTANDING WHICH METABOLITES
AND PROTEINS ARE ASSOCIATED WITH
CLINICAL OUTCOMES.
SO -- AND WE'VE STARTED TO DO
THAT WORK AND THEO BROMINE, I
BELIEVE WE'VE REPORTED TO BE
ADVERSELY ASSOCIATED WITH
CARDIOVASCULAR DISEASE, BUT I'LL
HAVE TO CHECK THE RESEARCH AND
GET BACK TO YOU ON THAT.
>> THANK YOU FOR THAT.
THERE'S ALSO A QUESTION THAT I
THINK YOU'VE ALREADY ANSWERED,
SOMEONE WAS ASKING ABOUT RACE
AND ETHNICITY BECAUSE OF BLP BLD
PRESSURE BEING HIGH.
YO.
>> I DIDN'T PRESENT THAT TODAY
BUT WE DO ANALYZE THE UNKNOWNS.
WE FOCUS ON THE NOANS BECAUSE
IT'S EASIER TO INTERPRET THEM,
THERE IS SOME INFORMATION IN THE
LITERATURE THAT CAN HELP US
UNDERSTAND THOSE FINDINGS, AND
ALSO IT'S HELPFUL TO LIMIT THE
NUMBER OF COMPARISONS IN TERMS
OF YOUR TYPE I ERROR, LIMITING
YOUR TYPE I ERROR.
WITBUT WE ALSO DO THE ANALYSIS F
UNKNOWNS AND IT'S PART OF THIS
PROCESS OF MOVING THE FIELD
FORWARD AND COMMUNICATE THOSE
RESULTS WITH THE LABORATORY THAT
GENERATED THE FINDINGS AND WE'RE
HAVING DISCUSSIONS ABOUT, YOU
KNOW, WHAT ARE HIGH PRIORITY
METABOLITES TO FURTHER
CHARACTERIZE.
THEY'LL USUALLY BE IN THE
SUPPLEMENT.
>> THANKS FOR THAT.
SOMEONE WANTS TO KNOW WHETHER
YOU HAVE DONE WORK ON
METABOLOMICS AND PROTEOMICS AS
IT RELATES TO THE MEDITERRANEAN
DIET.
THERE ARE SOME SIMILARITIES
BETWEEN THE TWO DIETARY
PATTERNS, SO --
>> YEAH, YEAH, SO IN THESE
ANALYSES THAT I SHOWED IN THE
ARIC STUDY, ATHEROSCLEROSIS IN
COMMUNITIES AND THE CRIC,
CHRONIC RENAL INSUFFICIENCY
COHORT STUDY, WE LOOKED AT OTHER
DIETARY PATTERNS THAT FOCUSED
TODAY ON THE DASH BUT WE ALSO
LOOKED AT THE MEDITERRANEAN DIET
SCORE AND REPORTED THOSE
FINDINGS.
SO SOME OF THEM OVERLAP WITH THE
DASH BECAUSE THEY'RE SIMILAR
DIETS AND SOME OF THEM ARE
UNIQUE, YOU KNOW, MANY OF THE
FISH METABOLITES COME OUT AS
REPRESENTATIVE OF MEDITERRANEAN
DIET, WHICH TRACKS.
THE PREDIMED INVESTIGATORS HAVE
ALSO DONE SOME METABOLOMICS
RESEARCH AND HAVE PUBLISHED
THOSE FINDINGS.
SO THOSE ARE GREAT, AND I THINK
IF THERE'S OTHER, YOU KNOW, WELL
CONTROLLED FEEDING TRIALS OF THE
MEDITERRANEAN DIET, IT WOULD BE
REALLY NICE TO EXPLORE MARKERS
OF THE MEDITERRANEAN DIET WITHIN
THAT SETTING.
>> VERY NICE.
SOMEONE WANTS TO KNOW WHETHER
THIS IS BEING USED IN CLINICAL
PRACTICE.
I THINK WE HAVE A LONG WAY TO
GO, BUT THE METABOLOMICS AND
PROTEOMICS BEFORE WE CAN START
USING THEM IN CLINICAL PRACTICE,
BUT I WONDER IF YOU CAN COMMENT
ON THAT.
>> I THINK SOME PEOPLE WOULD
LIKE TO DO METABOLOMICS AND
PROTEOMICS ON EVERYBODY,
INCLUDING IN CLINICAL PRACTICE
AND THERE'S A MODEL FOR DOING
THAT.
I AM OF THE VIEWPOINT THAT WE
NEED TO DEVELOP QUANTITATIVE
ASSAYS THAT MEASURE ABSOLUTE
INTAKE OR ABSOLUTE LEVELS IN THE
BODY, AND NARROW IT DOWN FROM
THOUSANDS OR HUNDREDS TO NOT A
SINGLE METABOLITE THAT
REPRESENTS A DIETARY PATTERN BUT
A SMALLER SUBSET OF METABOLITES.
SO I DON'T ENVISION MAYBE
REPEATING UNTARGETED
METABOLOMICS AND PROTEOMICS IN
EVERYONE WHO WALKS INTO THE
OFFICE, BUT EVENTUALLY, I THINK,
YOU KNOW, THAT'S THE IDEA, THAT
THESE BIOMARKERS COULD BE USED
IN CLINICAL SETTINGS.
>> VERY GOOD.
THANK YOU SO MUCH.
I THINK WE HAVE -- WE ARE AT THE
END OF OUR SESSION, AND I WANT
TO THANK EVERYONE FOR COMING,
AND I THINK WE HAVE A SLIDE TO
SHOW FOR THE NEXT UPCOMING
MEETING SOMETIME IN MAY OF THIS
YEAR.
YES.
SO THANK YOU AGAIN FOR JOINING
US.
PLEASE STAY TUNED FOR OUR NEXT
ONPA SEMINAR, WHICH IS TITLED
CARDIOVASCULAR HEALTH IN THE
PERIPARTUM PERIOD, DRIVERS &
DETERMINANTS, COMING UP MAY 29TH
OF THIS YEAR AT 1:00 P.M., AND
IT WILL ALSO BE AVAILABLE FOR
THE PUBLIC ON THE NIH WEBSITE.
OR THE NIH VIDEOCAST.
SO THANK YOU ALL VERY MUCH.
THANK YOU, DR. CASEY REBHOLZ,
PARTICULARLY FOR PRESENTING
THIS, AND HAVE A GOOD AFTERNOO,
EVERYONE.
>> THANK YOU SO MUCH, DR. PRATT.