Curcumin Therapy to Treat Vascular Dysfunction in Children and Young Adults With ADPKD
Phase: PHASE4
Conditions: Polycystic Kidney, Autosomal Dominant
Sponsor: University of Colorado, Denver

 
Study Protocol and Statistical Analysis Plan  
Project Title: Curcumin Therapy to Treat Vascular Dysfunction in Children and Young Adults with ADPKD  
NCT Number: NCT02494141 
Version Date: 03/12/2020 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Protocol #:  15-0902 
Project Title: Curcumin Therapy to Treat Vascular Dysfunction in Children and Young Adults with ADPKD  
Principal Investigator: Kristen Nowak, Ph.D. 
Version Date: 03/12/2020 
 
Specific Aims 
 
Autosomal dominant polycystic kidney disease (ADPKD) is the most common life-threatening genetic disease, 
affecting more than 600,000 Americans 1,2.  Although often considered to be a disease of adults, 
complications of ADPKD begin in childhood 3.  While the hallmark of ADPKD is the development and 
continued growth of multiple renal cysts that result in ultimate loss of kidney function 4, the leading cause of 
death among affected patients is cardiovascular in nature 1,2.  As much as 80% of all cardiovascular diseases 
(CVD) are associated with dysfunction and disorders of arteries 5.  Two of the greatest contributors are 
vascular endothelial dysfunction, most commonly assessed as impaired endothelium-dependent dilation 
(EDD), and stiffening of the large elastic arteries (aorta and carotid arteries) 6.  Importantly, both measures 
are independent predictors of future cardiovascular events and mortality 7-10.   
 
Adults with ADPKD demonstrate impaired EDD 11-13 as well as large elastic artery stiffening 14 , even in the 
absence of hypertension.  The mechanisms by which arterial dysfunction occurs in ADPKD are incompletely 
understood, but appear to involve increased oxidative stress and inflammation 15-17.  My preliminary data in 
children/young adults with ADPKD support that arterial dysfunction develops very early in the course of the 
disease, as evidenced by impaired brachial artery flow-mediated dilation (FMDBA; a measure of EDD) and 
increased aortic pulse-wave velocity (aPWV; a measure of large elastic artery stiffness).  There are presently 
very limited treatment options for the prevention of cardiovascular disease in adults with ADPKD, thus, 
childhood/young adulthood may represent a critical therapeutic window.  
 
An increase in total kidney volume (TKV) precedes the decline in kidney function, which is typically delayed 
until the 4th decade of life in patients with ADPKD, and TKV is a prognostic biomarker of future kidney function 
decline 18.  Thus, a decrease in rate of TKV growth, as measured by magnetic resonance imaging (MRI) is 
an important indicator of kidney disease progression early in the course of ADPKD.         
 
Curcumin is a safe, naturally occurring polyphenol found in the Indian spice turmeric that has a unique ability 
to activate transcription of key antioxidants, suppress inflammation, and reduce proliferation.  Curcumin 
reduces arterial dysfunction in various rodent models of vascular dysfunction 19,20, and my preliminary data 
demonstrate that it also reduces age-associated vascular dysfunction in humans.  Importantly, curcumin also 
slows cyst growth in kidney cells in vitro 21 and in vivo 22.  Thus, curcumin is a promising nutraceutical to slow 
progression of renal and cardiovascular disease in children with ADPKD.  However, there is presently no 
information available regarding the potential therapeutic benefits of curcumin to treat arterial dysfunction and 
slow kidney growth in this population.   
 
Accordingly, the primary aim of this application is to determine the efficacy of curcumin for improving FMDBA 
and reducing aPWV in 68 children/young adults with ADPKD.  A key secondary goal is to obtain insight into 
the mechanisms by which curcumin therapy may reduce arterial dysfunction.  Last, a third exploratory aim is 
to determine if treatment with curcumin slows kidney growth.  These aims will be tested in a double-blind, 
placebo-controlled, randomized clinical trial conducted in children/young adults aged 6-25 years with ADPKD. 
 
Hypothesis 1: Oral curcumin therapy will reduce vascular endothelial dysfunction (increase EDD) and large 
elastic artery stiffness in children and young adults with ADPKD as compared to placebo. 
 
Specific Aim 1: To determine non-invasively: a) FMDBA and b) aPWV before and after 12 months of oral 
curcumin (25 mg/kg/day) or placebo; and after 6 months in a sub-set of all local patients. 
 
Hypothesis 2: The improvements in arterial function observed with oral curcumin treatment will be associated 
with reduced oxidative stress and inflammation. 
 
Specific Aim 2: In a subset of patients aged 18-25 years, to measure aPWV and FMDBA during normal vs. 
reduced (via acute ascorbic acid infusion) oxidative stress, and to measure circulating/urine markers of 
oxidative stress and inflammation before and after 12 months of oral curcumin or placebo. 
 
Hypothesis 3: Oral curcumin therapy will reduce the rate of kidney growth in children and young adults with 
ADPKD as compared to placebo. 

 
Specific Aim 3: To quantify TKV measured with MRI before and after 12 months of oral curcumin or placebo. 
 
Impact on the field. This study has the potential to establish a novel, safe, and easy to deliver therapy for 
the treatment of arterial dysfunction, and possibly renal cystic disease, in an understudied population of 
children and young adults with ADPKD.  
 
Research Strategy 
 
Significance 
 
Autosomal Dominant Polycystic Kidney Disease (ADPKD) and Cardiovascular Disease (CVD).  ADPKD 
is the most common lethal genetic kidney disorder 1,2.   While the hallmark of ADPKD is the development and 
continued growth of multiple renal cysts that result in ultimate loss of kidney function 4, the leading cause of 
death among affected patients is cardiovascular in nature 1,2.  The proteins encoded by the PKD-1 and -2 
genes, polycystin-1 and -2, are expressed in vascular endothelial cells and smooth muscle cells of all major 
vessels, resulting in extrarenal manifestations of the disease 23.  There are presently very limited treatment 
options for the prevention of CVD in patients with ADPKD.   
 
ADPKD in Children and Young Adults.  Although often considered to be a disease of adults, ADPKD begins 
in childhood 3 and can in fact be diagnosed in utero 24.  Even in children with ADPKD, evidence of CV 
abnormalities begin to manifest, such as elevated blood pressure and increased left ventricular mass index 25-
27.  There are presently very limited treatment options for the prevention of CVD in adults with ADPKD.  Thus, 
childhood/early adulthood may represent a critical therapeutic window to prevent future CV complications. 
 
Vascular Function and CVD.  As much as 80% of all CVD is associated with dysfunction and disorders of 
arteries 5.  Two of the greatest contributors are the development of vascular endothelial dysfunction, most 
commonly assessed as impaired endothelium-dependent dilation (EDD), and stiffening of the large elastic 
arteries 6, both of which independently predict of future cardiovascular events and mortality 7-10. 
 
Vascular Function in ADPKD.  Adults with ADPKD demonstrate impaired EDD 11-13 as well as large elastic 
artery stiffening 14 .  Endothelial dysfunction occurs early in ADPKD and has been detected in normotensive 
adult patients with preserved kidney function 11-13.  My preliminary data in children and young adults support 
that vascular dysfunction develops very early in the disease, as evidenced by impaired brachial artery flow-
mediated dilation (FMDBA; a measure of EDD) and increased aortic pulse-wave velocity (aPWV; a 
measure of large elastic artery stiffness).  Targeting vascular dysfunction early in the course of ADPKD is more 
likely to alter the long-term course of the disease compared to later intervention, thus strategies to reduce 
vascular dysfunction in children and young adults with ADPKD have the potential for huge clinical impact.   
 
Integrative Physiological Mechanisms of ADPKD and Vascular Dysfunction.  The mechanisms by which 
vascular dysfunction develops in ADPKD are incompletely understood; however, reduced nitric oxide (NO) 
bioavailability is a critical contributor 12,17 and a common mechanism of both impaired EDD and increased 
arterial stiffness 6,28.  In ADPKD, oxidative stress and inflammation are increased and contribute to the decline 
in NO bioavailability 15-17.  Oxidative stress is defined as excessive bioavailability of reactive oxygen species 
(ROS) relative to antioxidant defenses.  Physiological stimuli, including inflammatory signaling, promote 
oxidative enzyme systems to produce ROS, including superoxide anion (O2● -) 29.  Superoxide reduces 
bioavailability of nitric oxide (NO) by a) reacting directly with NO to form peroxynitrite (ONOO-), another ROS; 
or b) oxidizing the essential cofactor (tetrahydrobiopterin) for NO synthesis by endothelial nitric oxide synthase 
(eNOS) 30,31.  Of note, plasma levels of asymmetric dimethylarginine (ADMA), an important inhibitor of eNOS, 
are increased in adult ADPKD patients with preserved kidney function 17.  Inflammatory signaling can also 
directly reduce NO bioavailability by down-regulating eNOS activity 32.  In turn, increased ROS stimulate pro-
inflammatory gene transcription and protein expression via the redox-sensitive pro-inflammatory transcription 
factor nuclear factor κ B (NFκB), thus further promoting a pro-inflammatory cascade in a vicious cycle 33,34.  
 
Total Kidney Volume in ADPKD.  An increase in total kidney volume (TKV) precedes the decline in kidney 
function, which is typically delayed until the 4th decade of life in patients with ADPKD, and TKV is a prognostic 
biomarker of future renal insufficiency 18.  TKV is a critical marker of disease progression in children with 
ADPKD, as renal function remains normal in nearly all affected children despite marked changes in kidney 
structure 3.  Thus, a decrease in rate of TKV growth, as measured by magnetic resonance imaging (MRI) is 
an important indicator of renal disease progression early in the course of ADPKD.         
 
Therapeutic Potential of Curcumin.  Curcumin, from the plant curcuma longa, is a safe, naturally occurring 
polyphenol found in the Indian spice turmeric that has long been used in traditional Indian medicine (see below 

for safety information).  In vivo, supplementation with curcumin improves physiological function in animal 
models of various diseases 35-37.  Although the exact mechanism of action is unknown, curcumin suppresses 
both oxidative stress and inflammation in vitro and in vivo 38-42.  Consequently, curcumin has been proposed as 
a safe and novel nutraceutical for intervention in several disease states 43. 
 
Curcumin and Vascular Dysfunction.  Curcumin administration reduces vascular dysfunction (increases 
EDD and/or reduces arterial stiffness) in rodent models of hypertension 20, diabetes 36,37, and aging 42.  These 
improvements are associated with increased vascular NO bioavailability and arterial eNOS expression, 
reduced vascular oxidative stress (decreased ROS, reduced oxidative damage, and increased antioxidant 
enzymes), and increased activity of hemoxygenase-1, which can promote anti-inflammatory pathways 20,36,37,42.  
In addition, my preliminary data support that curcumin also reduces vascular dysfunction associated with 
aging in healthy humans, consistent with published FMDBA data in post-menopausal women 44.  However, 
there is presently no information available regarding the potential therapeutic benefits of curcumin to treat 
vascular dysfunction in ADPKD, including children and young adults.  
 
Curcumin and Total Kidney Volume.  Curcumin has been of interest in nephrology for over a decade as a 
potential treatment of acute kidney injury (AKI) or chronic renal impairment in various rodent models of 
disease.  In addition to reducing histological evidence of kidney damage 45-48, its use in these models is 
associated with reduced evidence of oxidative stress and increased antioxidant activity 46,49,50, as well as 
reduced activation of the pro-inflammatory transcription factor NFκB and its downstream target tumor necrosis 
factor-α (TNF-α), concomitant with an increase in the anti-inflammatory transcription factor peroxisome 
proliferator-activated receptor-γ (PPARγ) 48.  Notably, curcumin also slows cyst growth in vitro in a dose-
response manner, using both the Madin–Darby canine kidney (MDCK) cell cyst model and an embryonic 
kidney cyst model 21.  Similarly, in the Pkd-1 deletion mouse model, curcumin improves renal histology and 
reduces proliferative index, cystic index, and kidney weight (normalized to total body weight) 22.  Thus, 
curcumin may also decrease the rate of TKV growth in children and young adults with ADPKD.   
 
In summary, cardiovascular complications are the leading cause of death in adults with ADPKD and are 
characterized by vascular dysfunction, which occurs very early in the course of the disease.  Intervening in 
affected children and young adults may represent a critical therapeutic window for the prevention of future 
cardiovascular events and mortality.  Curcumin is a novel, safe, and easy to deliver therapy with the potential 
to treat arterial dysfunction (an independent predictor of future cardiovascular events and mortality), and 
possibly renal cystic disease, in an understudied population of children and young adults with ADPKD. 
 
Summary of Significance.  The biomedical significance of the proposed work includes: 
 
ADPKD is the most common life-threatening genetic disease; 
 
There is an urgent need to establish the efficacy of interventions that may minimize CVD risk in patients 
with ADPKD; 
 
Presently, there are very limited treatment options for the prevention of CVD in patients with ADPKD; 
 
Many interventions targeting traditional risk factors have not successfully reduced CVD risk in patients 
with ADPKD, despite their efficacy in the general population; 
 
Childhood and young adulthood may be a critical therapeutic window for the treatment of arterial 
dysfunction and renal cystic disease in ADPKD; 
 
In a subsample of participants 18-25 yrs, insight will be provided into the integrative physiological 
mechanisms using novel translational research techniques that are non- to minimally-invasive. 
 
Innovation 
 
The innovative features of this application include:  
 
1. Intervening early in the course of ADPKD with treatment in children/young adults (6-25 years of age); 
2. Assessing two major contributors to arterial dysfunction and CVD risk in ADPKD: vascular endothelial 
dysfunction and large elastic stiffness; 
3. Determining the efficacy of a novel nutraceutical (curcumin) that is an inexpensive, safe, naturally 
occurring, and easy to deliver substance; 
4. In a sub-group of 18-25 year olds, using a translational approach to determine the physiological 
mechanisms contributing to any improvements in vascular dysfunction with curcumin, including: 
a. functional in vivo assessment of the role of oxidative stress in arterial dysfunction using acute 
antioxidant administration  
b. measurement of urine/circulating markers of oxidative stress and inflammation, utilizing state of 

the art mass spectrometry methods 
5. Gaining exploratory evidence of the efficacy of curcumin to slow kidney growth, which will provide the 
basis for a future R01 application employing a longer treatment duration and larger sample size. 
 
Approach   
 
Subjects.  After obtaining their written informed consent (ages 18-25) or assent (ages 6-17; plus parental 
consent), children and young adults aged 6-25 years with a diagnosis of ADPKD, based on the presence of  
bilateral renal cysts when there is a positive family history 51, and normal renal function with an estimated 
glomerular filtration rate (eGFR) >80 mL/min/1.73 m2 (using CKid Schwartz bedside equation for ages 6-17 52 
and the CKD-EPI equation for ages 18-25 53) will serve as subjects.  Patients will undergo screening and 
vascular testing at the University of Colorado Denver Division of Renal Diseases and Hypertension Clinical 
Vascular Physiology Laboratory, which is part of the Clinical Research Unit.  A local contracted laboratory 
(Quest) and medical records will be used for screening for out of state participants.  The University of Colorado 
Denver ADPKD Center has successfully employed this approach in previous and ongoing ADPKD clinical 
studies 54,55, including a recently completed trial by the co-mentor (Dr. Cadnapaphornchai) in 110 children and 
young adults with ADPKD 56.  Laboratory analysis will be performed at the University of Colorado Hospital or 
by Quest, and mass spectrometry analyses will be performed in the iC42 Clinical Research & Development 
Center.  Renal magnetic resonance imaging (MRI) scans will be performed at the Brain Imaging Center.   
Major inclusion/exclusion criteria are presented in the table below (Table 1).  Those who drop-out/do not 
complete the study will be encouraged to undergo an end-of study visit which will include identical 
measurements to the ones planned for session 5. 
 
 
 
Table 1       Inclusion Criteria 
Exclusion Criteria 
 Aged 6-25 years 
 ADPKD diagnosis based on bilateral renal cysts 
+ positive family history 51 
 Normal renal function with an estimated 
glomerular filtration rate (eGFR) 
>80 mL/min/1.73 m2 (using CKid Schwartz 
bedside equation for ages 6-17 52 and the CKD-
EPI equation for ages 18-25 53) 
 Ability to provide informed consent or assent 
 Currently taking a curcumin supplement  
 Current smokers or history of smoking in the past 12 months 
 Marijuana use within 2 weeks prior to FMDBA and aPWV testing 
 Antioxidant and/or omega-3 fatty acid use within the past 4 weeks prior to 
FMDBA and aPWV testing and for the duration of the study 
 Alcohol dependence or abuse 
 History of hospitalization within the last 3 months 
 Active infection or antibiotic therapy 
 Pregnancy, lactation, or unwillingness to use adequate birth control 
 Body-mass index >95th percentile in ages 6-17 or >40 kg/m2 in ages 18-25 
(vascular measurements can be inaccurate if severely obese) 
 Inability to cooperate with/clinical contraindication for MRI including severe 
claustrophobia, implants, devices, or non-removable body piercings  
 
Experimental Design (Figure 1).  A 1-year randomized, placebo-controlled, double-blind design study with 
curcumin will be conducted.  Subjects will undergo telephone and laboratory screening for inclusion/exclusion 
criteria and will then be randomly assigned to either curcumin or placebo.  Members of the investigative team 
involved in the acquisition and analysis of data will be blinded to the treatment status.  Vascular function 
measurements will be made under supine, overnight fasted (water only) conditions before and after 1-year of 
the intervention, and at 6-months in local patients to reduce variability over time and better establish timeline of 
change.  Females who are post-pubertal will be scheduled during the first week of their menstrual cycle, when 
estrogen and progesterone are lowest, minimizing the impact of hormonal variations on vascular function 57. 
 
 
Figure 1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Session 5 
Mid-study Measurements            
(local patients only)  
Subjects randomly assigned to curcumin or placebo 
Preliminary screening by telephone; laboratory screening 
Baseline Measurements 
Baseline Measurements 
6 months curcumin 
Mid-study Measurements            
(local patients only)  
6 months placebo 
Post Measurements 
Post Measurements 
6 months placebo 
6 months curcumin 
Session 1  
Sessions 2 & 3 
Sessions 6 & 7 
Session 4 

In state subjects will come to the Clinical Vascular Physiology Laboratory (sessions 1, 2, 4, 5, and 6) or the 
Brain Imaging Center (sessions 3 & 7) on 7 separate occasions.  Out of state participants (approximately half) 
will travel twice to Colorado. Sessions 2 & 3 and then 6 & 7 will occur on consecutive days during the same trip 
(for a total of 2 out of state trips during the study).  Session 1 (and subsequent safety labs) will be 
arranged at a local contracted laboratory (Quest) for out of state participants.   
 
 
Session 1: Screening measurements  
 
Session 2: Baseline measurements 
 
Vital signs 
 
Blood sampling for measurement of circulating curcumin levels, serum creatinine, ALT and AST 
and urine sampling for markers of oxidative stress (all subjects); blood sampling for markers of 
inflammation (subset of subjects 18-25 years)  
 
FMDBA (EDD) and aPWV (arterial stiffness) (all subjects), with and without acute inhibition of 
oxidative stress via systemic i.v. infusion of ascorbic acid (subset of subjects 18-25 years) 
 
Endothelium-independent dilation to sublingual nitroglycerin (subset of subjects 18-25 years) 
 
Session 3: Baseline measurements 
 
Abdominal MRI for subsequent analysis of TKV (all subjects) 
 
Begin study treatment (curcumin or placebo) 
 
Session 4: Safety lab check  
 
Session 5: Mid-study measurement of FMDBA and aPWV for local (in state) patients only; safety lab 
check (all patients)  
 
Sessions 6 & 7 (month 12): Identical to Sessions 2 & 3 (note: subjects will not take a dose the a.m. 
prior to testing session to avoid any acute effects) 
 
Curcumin.  Curcumin will be delivered as a dose of 25 mg/kg/day rounded to the nearest 100 mg, delivered as 
powder measured in 100 mg and 500 mg scoops (LongvidaTM, Verdure Sciences).  The placebo will contain 
inert substances (soy lecithin, stearic acid, ascorbyl palmitate, carrot extract) of similar proportions and 
amounts as in the active powder.  The powder delivery will improve adherence, particularly among young 
children who may have difficulty swallowing capsules.   The powder will be delivered mixed with a choice of 
pudding yogurt, or other foods of similar consistency.  Absorption of LongvidaTM is the same when taken with a 
small amount of food compared with water or milk.   
 
Curcumin has been investigated extensively for safety and toxicity because of its use in foods.  Toxicology 
studies have been performed on the effects of curcumin in rodents and humans that are summarized below in 
the overall context of the present study.   
 
Background.  Curcumin is the active ingredient in the Indian spice turmeric and gives it a yellowish color.  It is 
found naturally in the root of the plant Curcuma longa and has traditionally been used for the prevention and/or 
treatment of ailments related to the skin, liver, gastrointestinal tract, and the common cold 58.  Epidemiological 
studies have shown that societies that regularly consume curcumin in their diet have markedly reduced 
mortality due to coronary heart disease 59.  More recently, curcumin’s potential benefits have been assessed in 
other pathologies such as cancer, arthritis, Alzheimer’s disease and hypercholesterolemia.  In addition, 
curcumin has been shown to attenuate oxidative stress and inflammation both in vitro and in vivo 38,39,41,42,60. 
 
History/Testing.  Because of its color, flavor and antioxidant-stabilizing abilities curcumin is commonly used in 
yellow mustard, pickles and sauces.  Curcumin has been used to flavor foods and treat ailments for over 5,000 
years and is generally recognized as safe (GRAS) by the FDA (this is also true specifically of LongvidaTM).  
Studies conducted in rats and humans using standard toxicology protocols have shown no toxic effects of 
curcumin in vivo at doses as high as 8 grams/day (8,000 milligrams/day), i.e., several-fold higher than the 
dose proposed in the present application 61-64.   
 
Human Studies.  Few side-effects of curcumin have been reported with doses up to 8-12 grams/day (8,000-
12,000 milligrams/day) of supplementation in adults 61,62,65 and children 64,66,67.  The few side-effects reported 
primarily involve gastrointestinal discomfort 61,65,67.  
 
 
Improving Bioavailability: Choice of Compound.  To enhance the bioavailability of curcumin, manufacturers 
have developed formulations that may prevent glucuronidation, as unprocessed curcumin is insoluble in water 
and quickly metabolized by glucuronidation.  The compound proposed in the current application is a solid lipid 
curcumin particle (SLCP, LongvidaTM, Verdure Sciences) that increases plasma levels of curcumin significantly 
compared to traditional formulations in healthy volunteers 64 and is safe in both this group and osteosarcoma 

patients, including children 64.  SLCP supplementation in rats with 720 mg/kg body mass/day, equivalent to 40 
g/day (40,000 mg/day) for a 60 kg person, for 90 days had no adverse effects 63.  
 
In summary: 1) curcumin is a naturally occurring and widely used spice in southeastern Asian cultures and 
poses only minimal risks at very high doses; and 2) all of the safety studies performed in adults and children 
have used maximal doses of curcumin that are significantly higher than those proposed in the present 
application (detailed rationale regarding doses and duration of treatment is below). 
 
Rationale for Proposed Dose (25 mg/kg/day), Frequency (once a day) and Duration (12 months)  
Dose (25 mg/kg/day) 
 Dosing is normalized to body mass to account for wide differences across the included age range. 
 This normalized dose is based off a 2,000 mg dose delivered to an 80 kg adult, which is similar to published  
work of others 61,62,64,67 and well below maximal dosing levels.  This dose has also been recommended by 
Verdure Sciences, who is providing the SLCP powder. 
 Additionally, 25 mg/kg/day was selected because 2,000 mg/day has been consistently shown to be both 
safe and well tolerated in adults 61,62,64, and children 64,67, is unlikely to elicit gastrointestinal discomfort 61,67, 
and improved vascular function in preliminary studies performed in aging adults (see Preliminary Results). 
 While older children and young adults could have instead been prescribed to swallow capsules, all 
participants will ingest the curcumin as powder mixed with pudding or yogurt, to standardize intake across 
all ages. 
 
Dosing Frequency (once a day) 
 The curcumin (or placebo) powder will be administered once a day in the morning for standardization.   
 
Duration of Treatment (12 months) 
 Given my previous experience with interventions to improve arterial function in the context of healthy aging, 
I expect 12 months to be a sufficient duration 68,69.  While I recognized that the duration is likely too short for 
a significant decrease in rate of TKV growth, any trend from this exploratory aim will be useful to design a 
future, longer duration R01 grant application. 
 Although my preliminary data (see below) support that curcumin has beneficial effects in healthy aging on 
FMDBA at 3 months, a 12 month treatment period was selected because 1) ADPKD may have a more 
severe effect on vascular function than healthy aging; 2) reversal of structural properties of the arterial wall 
associated with arterial stiffness may require a longer duration of treatment; 3) this allows for exploratory 
insight into changes in TKV, which is supported by cell and animal data 21,22. 
 I considered including multiple groups with different outcomes, including a shorter treatment length to 
evaluate vascular function; however, this design would greatly reduce power and further increase cost of the 
study.  Instead, I opted to add evaluation vascular function in local patients (~50% of the total n) at 6 
months, in addition to 12 months, in order to account for variations in adherence and changes in 
development over time (i.e. puberty) that may increase variability. 
 Importantly, I recognize that aPWV changes across puberty 70-72, whereas the available data suggest that 
FMDBA does not 73.  However, this should not limit the interpretation of the results as 1) randomization will 
ensure similar changes across the active and control groups; 2) puberty will be accounted for in statistical 
analysis; and 3) vascular measurements at 6 months in local patients will help account for such changes.        
 Curcumin has been tested extensively and used in the food/manufacturing industry.  It is a natural dietary 
component, and our proposed dose has been repeatedly shown to be well-tolerated and elicit no side 
effects.  Thus, there is no evidence to suggest the possibility of adverse effects associated with the 
proposed 12 months of increased intake of curcumin.   
 
Study Modifications to Account for COVID-19 Outbreak.  On account of COVID-19 and Institutional policies 
in place regarding work-related travel, the length of treatment of participants in may be extended due to 
extenuating circumstances.  The end-of-study visit may be delayed due to concerns over air travel, in which 
case participants will be asked to continue taking the curcumin/placebo, as they have been doing for the past 
year, and we will reschedule their end-of-study visit at the earliest time point that seems appropriate (hopefully 
within a couple of months).  We will obtain in-person data at this time, but will ask participants to go to our 
contract laboratory for the clinical labs at the planned 12 month time point if possible. We do not see any 
additional risk posed to participants by extending their treatment period, as curcumin already has a very 
favorable safety profile.   
 

Summary of risks associated with curcumin.  Based on preliminary studies (see below) and an extensive 
review of literature, the proposed doses of curcumin are both physiologically relevant and predicted to be well-
tolerated by the great majority of study participants.  The primary risk associated with curcumin 
supplementation is the possibility of temporary/transitory gastrointestinal distress (flatulence, constipation, 
diarrhea, nausea) in a very small percentage of individuals 61,65,67.  
 
Measurements. 
 
Screening Measures.  Subjects will undergo a medical history and physical exam, including growth 
parameters and Tanner staging 74-76, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and 
a pregnancy test for all female participants of possible childbearing potential (Tanner Stage 2 or higher).  
Serum creatinine will be measured for determination of eGFR using the CKid   Schwartz bedside equation for 
ages 6-17 52 and the CKD-EPI equation for ages 18-25 53.  A local contracted laboratory (Quest) and medical 
records will be used for out of state participants.  EMLA cream may be used as needed to numb the skin prior 
to drawing blood.     
 
Measures of Adherence and Safety: 
 
Adherence.  As we expect ~50% of all participants to be from out of state, progress will be reviewed with a 
monthly phone inquiry of medication supply.  Please note, due to the long history and prestige of University of 
Colorado PKD Center studies, our families have tremendous relationships with our research staff (i.e. 
investigators and research coordinators).  The co-mentor, Dr. Cadnapaphornchai, has successfully employed 
this approach in previous clinical trials in children/young adults with ADPKD that have included ~50% out of 
state patients 54,56.  We expect the minimal side-effects associated with curcumin to favor adherence.  
Participants will have powder mailed every 3 months and will also be asked to mail back the bottle for a volume 
check.   
 
Safety Checks.  While adverse events are not anticipated, a safety questionnaire will be administered over the 
phone monthly.  Serum creatinine, ALT and AST will be measured after 1, 6, and 12 months, and for those out 
of state, will be performed at a local contracted laboratory (Quest).  Home pregnancy tests will be required 
monthly for all girls of possible childbearing potential (>Tanner Stage 2).  All subjects will be given a digital 
blood pressure monitor (A&D Medical, UA767) with an appropriately sized cuff and the subject/parent will be 
instructed in its use.  Home blood pressures will be taken monthly and reported during phone safety checks.  
This approach has been successfully employed previously by the co-mentor, Dr. Cadnapaphornchai 54,56.    
 
Plasma Curcumin Concentrations will be assessed at baseline and 12 months by liquid chromatography-
tandem mass spectrometry (LC-MS/MS) at the University of Rhode Island 77.  Additionally, curcumin 
concentrations will be measured by the in red blood cells, the buffy coat, and plasma with the addition of citrate 
to prevent the degradation of metabolites.   
 
Randomization will be performed by the statistician, using a blocked randomization sequence, with 
stratification by age group (6-13 or 14-25 yrs). 
 
Outcome Measures.  The following measurements will be made at baseline and after 12-months of treatment: 
 
Primary (Functional) Outcomes (Specific Aim 1)  
 
Brachial Artery FMD and Endothelium-Independent Dilation.  FMDBA will be determined using high-
resolution ultrasonography (Toshiba Xario 200) as described originally by Celermajer et al. 78 and used 
recently by the applicant 68,79,80.  FMDBA is a widely used, non-invasive method inducing reactive hyperemia to 
assess endothelium-dependent dilation.  It is validated in children 78,81 and is mainly dependent upon NO 
production 82,83.  Brachial artery endothelium-independent dilation will be determined in the sub-group of 
participants aged 18-25 yrs by measuring brachial artery dilation for 10 minutes after administration of 
sublingual nitroglycerin (0.4 mg) 67,84,85.  This assessment is key to the interpretation that any improvements in 
dilation are indeed dependent upon endothelial production of NO rather than improved smooth muscle 
relaxation to NO 86.  The use of nitroglycerin is currently safely employed in an NIH-funded, IRB-approved 
protocol in adults with ADPKD at the University of Colorado Denver.  A commercially available software 
package (Vascular Analysis Tools 5.8.1, Medical Imaging Applications) will be used to acquire and analyze 
ECG-gated brachial artery diameters.  Doppler blood flow velocity will be obtained for estimation of shear rate, 
which will be entered as a covariate if indicated 87-89. 
 
Aortic Pulse-Wave Velocity.  aPWV will be determined as described in detail previously by applicant 69,90 and 
validated in children 71,91.  Briefly, a transcutaneous custom tonometer (Noninvasive Hemodynamics 

Workstation, Cardiovascular Engineering Inc.) will be positioned at the carotid and femoral arteries to non-
invasively assess aPWV, as distance/time between the foot of the arterial waveforms 92.  As a secondary index 
of arterial stiffness, ultrasound imaging of the carotid artery will also be performed for calculation of carotid 
artery compliance and the β-stiffness index (in conjunction with tonometry), as described previously 96,97.  The 
images will also be assessed for measurement of carotid artery intimal-medial thickness (IMT), which is used 
in the calculation of carotid artery compliance and the ß-stiffness index. 
 
Outcomes for Specific Aims 2 (Only in the sub-group aged 18-25 years (except for urinary markers)) 
 
Oxidative Stress-Associated Suppression of EDD and Large Elastic Artery Stiffness.  The influence of 
oxidative stress on FMDBA and aPWV (and carotid artery compliance) will be determined by infusing a 
supraphysiological dose of ascorbic acid (American Regent Labs) known to scavenge superoxide or 
isovolumic saline, as used previously by the applicant 68,93, and others 84,94-96.  This procedure is currently 
safely employed in an NIH-funded, IRB-approved protocol in adults with ADPKD at the University of Colorado 
Denver.  The infusion will be performed only if cannulation in a vein on the dorsal aspect of the contra-lateral 
hand is successful.  0.075 g ascorbic acid/kg fat-free mass dissolved in 150 mL of saline will be infused 
intravenously at 5 mL/min for 20 min, followed immediately by a “drip-infusion” at 0.5 mL/min over 60 min.  
Vascular measurements will be made during the “drip infusion” when peak plasma concentrations of ascorbic 
acid occur 84.  The difference in FMDBA and aPWV (and carotid artery compliance) during ascorbic acid vs. 
saline infusion indicates the modulation of EDD and arterial stiffness by oxidative stress 84,94.  These measures 
will be performed in the sub-group (18-25 yrs) only. 
 
Markers of Oxidative Stress.  Urinary 8-iso-prostaglandin F2α (8-isoprostane) and 8-hydroxy 2 
deoxyguanosine (8-OHdG) will be measured by LC-MS/MS as a markers of lipid peroxidation 97 and oxidative 
DNA damage 98, respectively, as oxidative damage is increased ADPKD 17.  While Aim 2 is primarily in a sub-
group aged 18-25 yrs, these oxidative stress measurements will be performed in all participants (6-25 yrs), as 
urine can be collected non-invasively.   
 
Inflammatory Markers.  Plasma C-reactive protein (immunoturbidimetric method) and plasma interleukin-6 
(ELISA; R&D Systems) will be measured by the University of Colorado Hospital as circulating inflammatory 
markers 79,99, as both are increased in ADPKD 16.  To minimize the amount of blood collected in children <18 
yrs, these measures will only be performed in the sub-group aged 18-25 yrs.   
 
Outcome for Specific Aims 3 
 
MRI Measurement of TKV.  A 1.5 Tesla Visart System (Toshiba America Medical Systems) or comparable 
system will be used for all studies.  Renal images will be acquired in similar manner and volumetric 
measurements determined as described for the CRISP study 100.  No contrast agents will be utilized for the 
study.  The radiologist, MRI study technicians and the applicant, who will calculate TKV, will be blinded 
regarding group assignment.  For analysis, DICOM images will be de-identified and evaluated by a single 
analyst (the applicant) using Analyze software (Analyze 9.0, Mayo Foundation, Rochester, MN).  To account 
for normal growth in children, TKV will be adjusted for height 56.  Feasibility of this aim is supported by the fact 
that the co-mentor, Dr. Cadnapaphornchai, recently completed an intervention study with MRI measurement of 
TKV in 110 pediatric and young adult patients with ADPKD 56.  In addition renal parenchyma (TKV – cyst 
volume) will be measured. 
 
Additional Blood Samples 
For subjects signing the Informed Consent Addendum, an additional 5 mL of serum and 9 mL of plasma in 
adults and an additional 1 mL of serum and 2 mL of plasma in children will be collected for future research.  In 
addition, 20 mL of whole blood will be collected from adult participants only.  PBMCs will be isolated from 
heparinized whole blood using Histopaque-1077 as described previously 106,107.  To provide mechanistic insight 
into biological pathways that may be altered by curcumin (for example, mammalian target of rapamycin 
(mTOR)-S6 kinase (S6K) signaling), the PBMCs will be washed and protein will be extracted for Western blot 
analysis.    
 
Expected Results.  In children/young adults with ADPKD, 12 months of curcumin compared to placebo will: 
 
 
Hypothesis 1:  
 ↑ FMDBA (↑ vascular EDD) 
 ↓ aPWV (↓ large elastic artery stiffness) 
 
Hypothesis 2 (Only in participants 18-25 years of age): 

● Have little or no effect on FMDBA and aPWV in the presence of ascorbic acid, whereas ascorbic acid is 
expected to significantly improve both of these measures in placebo-treated subjects, indicating a 
curcumin-mediated reduction in oxidative stress-related suppression of vascular function 
● ↓ urinary 8-isoprostane and 8-OHdG, indicating reduced systemic oxidative stress 
● ↓ plasma C-reactive protein and interleukin-6, indicating reduced systemic inflammation 
 
Hypothesis 3 
 Tend to ↓ the progression of TKV (adjusted for height), which can be examined further in a longer-
duration, larger sample size, future R01 grant  
 
Power Calculations and Statistical Analysis.  Power calculations are based on clinically meaningful 
differences in the primary outcomes (FMDBA and aPWV), limited published literature using curcumin in a 
healthy older population 44, and experience of the applicant regarding the variability of the proposed outcomes 
68,69,79.  A sample size of 27 in each group with have 90% power to detect a mean increase in FMDBA of 1.5%, 
given a standard deviation of 2.0 68, with α = 0.025, adjusted for two primary endpoints.  Akazawa et al. 44 
demonstrated in healthy post-menopausal women a ∆FMDBA of +1.5% with curcumin compared to a decrease 
in FMDBA of -0.2% in controls.  Similarly, a sample size of 27/group will have 90% power to detect a mean 
decrease of aPWV of 100 cm/sec given a standard deviation of 125 69 with α = 0.025, adjusted for two primary 
endpoints.  To account for a potential dropout of 20%, 34 subjects/group will be enrolled.    
 
A sub-sample size of 10 in each group (of 18-25 year olds) will have 89% power to detect a 50% reduction in 
∆FMDBA with ascorbic acid (given ∆FMD = +3.0+1.0% with ascorbic acid in the placebo group 68), with a two-
sided α = 0.05.  Aim 3 will be considered exploratory, thus power was not calculated.     
  
Differences between groups at baseline will be assessed by an independent t-test, and a linear mixed-effects 
model will be fit to compare curcumin vs. placebo (independent variable) with respect to change in outcomes at 
6 and 12 months (dependent variables), with covariates and potential confounders (including puberty) 
incorporated into the model.  This approach will account for the use of multiple time points and is flexible to 
missing (at random) data.  Although I do not expect different responses based on sex or race/ethnicity, and the 
study is not powered to assess these factors, stratified analysis will be performed and any trends will be noted 
and studied in a larger follow-up study.  Dropouts will be handled using an intent-to-treat analysis. 
 
Study Time Line.  I will use the initial 8 weeks to begin subject recruitment and screening.  The first subjects 
will begin treatment with curcumin or placebo in month 3 of the proposed overall 5-year funding period.  The 
last patients enrolled will complete the study by approximately month 54.  Vascular and MRI measurements 
will be performed over this period.  Mass spectrometry assays will be performed annually to detect any 
problems early in the course of the study.  Statistical analysis and manuscript preparation will be undertaken 
during the last 6 months (months 54-60).  This is an appropriate rate of subject enrollment and experimental 
testing based on previously completed intervention studies in this population (which also included ~50% out of 
state patients) by the co-mentor Dr. Cadnapaphornchai 54,56. 
 
Recruitment Plans.  Since 1985, the University of Colorado Denver has maintained an ADPKD registry and  
mailing list.  Included in this registry are children and young adult ADPKD patients from the Denver Metro Area 
and all over the United States.  In addition to the University of Colorado Denver registry, I will advertise through 
the national PKD Foundation.  The ADPKD center at the University of Colorado Denver has longstanding 
expertise and national recognition, and therefore, is able to attract ADPKD patients from all over the United 
States for clinical studies.  Of note, the University of Colorado Denver is currently the only group in the world 
performing research studies in children with ADPKD.  The feasibility of recruitment is supported by the recently 
completed intervention study performed by the co-mentor Dr. Cadnapaphornchai, which included 110 pediatric 
and young adult patients with ADPKD (including ~50% out of state) followed for a three-year period 56, and 
these participants will be recruited for the proposed research study.   
 
Potential Problems, Alternative Strategies, and Future Directions.  Although subject recruitment and 
retention always are challenges, I am confident in my ability to complete the study in the proposed timeline 
given our location in the densely populated Denver-Boulder metro area and our department’s 30 years of 
experience in studying patients from around the country with ADPKD, including children and young adults.  I 
should have few difficulties with the proposed experimental procedures and protocols as either a) I have 
expertise in the techniques in other populations; or b) the methodology is already established within our 
ADPKD research group, and I will be trained to perform these new skills by my mentorship team. 
 
I recognize that alternative approaches exist for examining the effects of curcumin in children and young adults 
with ADPKD.  For example, one alternative would be to undertake a larger study to examine harder 

cardiovascular endpoints than the intermediate outcomes proposed in the current application.  However, this 
approach requires very large sample sizes, and the validated endpoints of FMDBA and aPWV will provide initial 
insight that can guide a future larger-scale clinical trial using hard clinical endpoints.  Similarly, I acknowledge 
that long-term follow-up is required to determine if curcumin has beneficial effects into later adulthood, and this 
is also a potential future direction following this initial study.  I recognize that the proposed study duration and 
sample size will only provide a trend for any changes in TKV with curcumin, and such exploratory evidence will 
provide the basis for a future R01 application employing a larger sample size and treatment duration.   
 
Last, I recognize the possibility of negative data; however, such data are still important, as a recent state of the 
art review on prospective and novel treatments of ADPKD identified curcumin as a potential therapy 101, and 
either positive or negative results are critical in continuing to advance treatment options.  Importantly, the 
proposed Career Development plan will provide the scientific and professional skills required to conduct future 
randomized controlled trials in this patient population, and I will continue to pursue other novel interventions 
through small pilots and in conjunction with my collaborators performing pre-clinical research throughout the 
award’s duration.  In the event the study is negative, other potential therapies for a future R01 application 
include the mitochondria-targeted antioxidant MitoQ and metformin.  My training in epidemiology will also allow 
me to identify other targets for future clinical trials.  In addition, I will have a unique dataset of longitudinal 
changes in vascular function, oxidative stress, and TKV, which may generate novel hypotheses.              
 
Preliminary Results: all data are presented as mean+s.e. 
 
I. Vascular Endothelial Dysfunction in Children and Young Adults  
with ADPKD 
 
I assessed FMDBA in 9 children/young adults (ages 6-25 yrs) with ADPKD and normal kidney function, and 9  
age- and sex-matched healthy controls (16+2 years; 1M/8F).  Compared to 
the healthy control group, FMDBA was impaired (Figure 2) in the children and 
young adults with ADPKD, indicating vascular dysfunction very early in the 
course of the disease.  Baseline brachial artery diameter (3.1+0.09 [ADPKD] 
vs. 3.1+0.16 cm [control]) did not differ between groups.  Blood pressure 
(108+13/59+7 vs. 109+5/59+2 mmHg) and body-mass index (22.0+2.9 vs. 
21.6+1.4 kg/m2) also did not differ between groups.   
 
II. Large Elastic Artery Stiffness in Children and Young Adults  
with ADPKD 
 
I also assessed aPWV in the same 9 children 
and young adults with ADPKD and the same 
9 age- and sex-matched healthy controls.  
Compared to the control group, aPWV was 
greater (Figure 3), indicating increased large-
elastic artery stiffness even at a young age.  
Carotid artery β-stiffness index, a measure of 
local large-elastic artery stiffness, was also 
greater in the ADPKD group (Figure 4).  
Resting heart rate did not differ between groups (66+8 [ADPKD] vs. 63+2 bpm [control]).  
 
III. Curcumin to Treat Vascular Dysfunction 
 
My graduate laboratory has assessed the efficacy of curcumin therapy 
(same company and similar normalized dose as proposed: 2,000 mg/day, 
LongvidaTM, Verdure Sciences) to reduce age-associated vascular 
dysfunction.  Compared to placebo, FMDBA was improved (Figure 5) in 5 
healthy adults (65+3 years, 2M/3F) treated for 3 months with curcumin, as 
compared to placebo.  The curcumin was well tolerated in all participants 
without any adverse effects.  As aging is associated with impaired FMDBA 
similar to ADPKD, curcumin may also reduce vascular dysfunction in  
children/young adults with ADPKD.  Please note, aPWV and B-stiffness index were not assessed in this 
preliminary study, as it may require more time to change in response to an intervention.  
 
Human Subjects 
 
Figure 2 
Figure 5 
Figure 3 
 
Figure 4 

1. Risks to the Subjects 
 
a) Human Subjects Involvement and Characteristics. 
 
Subjects.  After obtaining their written informed consent (ages 18-25) or assent (ages 6-17; plus parental 
consent), children and young adults aged 6-25 years with a diagnosis of autosomal dominant polycystic kidney 
disease (ADPKD), based on the presence of bilateral renal cysts when there is a positive family history 51, and 
normal renal function with an estimated glomerular filtration rate (eGFR) >80 mL/min/1.73 m2 (using CKid 
Schwartz bedside equation for ages 6-17 52 and the CKD-EPI equation for ages 18-25 53) will serve as subjects 
will serve as subjects.  Patients will undergo screening and vascular testing at the University of Colorado 
Division of Renal Diseases and Hypertension Clinical Vascular Physiology Laboratory.  A local contracted 
laboratory (Quest) and medical records will be used for screening for out of state participants.  The University 
of Colorado Denver ADPKD Center has successfully employed this approach in previous and ongoing ADPKD 
clinical studies 54,55, including a recently completed trial by the co-mentor (Dr. Cadnapaphornchai) in 110 
children and young adults with ADPKD 56.   Laboratory analysis will be performed at the University of Colorado 
Hospital, and mass spectrometry analyses will be performed in the iC42 Clinical Research & Development 
Center.  Renal magnetic resonance imaging (MRI) scans will be performed at the Brain Imaging Center. 
 
To be eligible to participate in this research, volunteers must meet the following criteria:  
 
Inclusion Criteria: 
 
 
1) Aged 6-25 years (rationale: 6 was chosen as the lower age limit, as children younger than 6 may have 
difficulties remaining still for the vascular and MRI measurements; 25 is the upper limit because the focus 
of the study is in on children and young adults) 
 
2) ADPKD diagnosis of ADPKD based on the presence of bilateral renal cysts when there is a positive family 
history 51 
 
3) Normal renal function with an estimated glomerular filtration rate (eGFR) >80 mL/min/1.73 m2 (using CKid 
Schwartz bedside equation for ages 6-17 52 and the CKD-EPI equation for ages 18-25 53) 
 
4) Ability to provide informed consent or assent 
 
 
Exclusion Criteria: 
 
 
1) Currently taking a curcumin supplement  
 
 
2) Current smoking or history of smoking in the past 12 months (rationale: changes in smoking habits during 
study involvement could confound the results) 102 
3)   Marijuana use within 2 weeks prior to FMDBA and aPWV testing (rationale: use may influence vascular 
function and cardiovascular changes, including blood pressure and heart rate) 
4)   Antioxidant and/or omega-3 fatty acid use within the past 4 weeks prior to FMDBA and aPWV testing and 
for the duration of the study (rationale: use may confound the effect of the acute ascorbic acid infusion on 
vascular function) 
5)   Alcohol dependence and abuse 
 
6) History of hospitalizations within the last 3 months 
 
7) Active infection or antibiotic therapy 
 
8) Pregnancy, lactation, or unwillingness to use adequate birth control.  All female participants of possible 
childbearing potential (Tanner Stage 2 or higher) will be required to take a pregnancy test at initiation of the 
study and then monthly during the study.  
 
9) Body-mass index >95th percentile using sex, age, and height for children 6-17 years of age, or > 40 kg/m2 
for young adults 18-25 years of age (rationale: vascular measurements can be inaccurate in severely 
obese patients).  The PI and co_I’s may use discretion to allow inclusion of children with body-mass index 
>95% if vascular measurements seem reasonable to perform.   
 
10) Inability to cooperate with or clinical contraindication for magnetic resonance imaging (MRI) including  
severe claustrophobia, implants, devices, or non-removable body piercings 
 
Special classes of subjects considered vulnerable populations will not be included in the study.  Dr. Chonchol 
will make final decisions on all patient eligibility.  
 

b) Sources of Research Materials.  This is a prospective study of newly recruited human subjects aged 6-25 
years with ADPKD.  The data collected will be used exclusively for research purposes.  All subject identities 
and records will remain strictly confidential. 
 
c) Potential Risks.  We see no psychological, social, or legal risks beyond those of participation in health-
related research in general.  The potential physical risks of participating in the proposed experiments are 
reasonably small.  The vascular measurements associated with Specific Aim 1 are completely non-invasive.  In 
children 6-17, the only invasive measurement with be collection of a small amount of blood.  All of the vascular 
procedures and the MRI scans have been used previously by the team of investigators in this patient 
population without complications.  Importantly, all of the procedures will be performed in University of Colorado 
Division of Renal Diseases and Hypertension Clinical Vascular Physiology Laboratory, a facility with on-site 
full-time supervision including nursing, safety equipment and established emergency procedures.  Moreover, 
the research will be overseen by a study-specific Data Safety Monitoring Board (DSMB) (described in more 
detail under Data and Safety Monitoring Plan). 
 
The risks associated with the experimental protocols include: 
Venous Catheter (ages 18-25 only) - Discomfort associated with insertion of the needle; local bleeding and 
a small hematoma (~10% of cases); risk of infection of a hematoma or significant external blood loss (<1 in 
1000), risk of fainting. 
 
Blood draw – Discomfort associated with the insertion of the needle; risk of bruising or feeling lightheaded 
or faint.  
 
Endothelium-Dependent Dilation (Brachial Artery Flow-Mediated Dilation [FMDBA]) - Inflating the blood 
pressure cuff during this procedure may cause a mild to moderate intensity “pins and needles or numbing” 
sensation that resolves when the cuff is deflated.     
 
Magnetic Resonance Imaging - The risk of performing abdominal MRI is minimal.  The magnetic field 
generated within the MRI is not harmful but can cause metal within the body to heat up or electronics to 
stop working.  All subjects will be questioned regarding the presence of metal or electronic devices inside 
their body.  All subjects with either metal implants or implanted electronic devices will be excluded from the 
study.  As the MRI tube is a small round tube, it may make subjects who experience claustrophobia 
uncomfortable, thus such individuals will be excluded.  The most common minor side effect of having an 
MRI exam is flashing lights in the eyes.  This is caused by the magnetic waves and is not harmful.  Some 
people also experience warmth and reddening of the skin, which usually goes away after a few minutes. 
For all female participants of possible childbearing potential, a negative pregnancy test prior to MRI will be 
required.  Severe claustrophobia is an exclusion criterion for the study.  Thus, undue stress due to a 
confined space during the MRI procedure will not be a potential risk.   
 
Medications/Dietary Supplements - With any medication used in testing, there is small risk of an allergic 
reaction.  Importantly, a physician will be available during the ascorbic acid infusion and nitroglycerin 
administration (ages 18-25 only) for assistance in the unlikely case of an adverse event.   
 
Reactions reported with the use of the specific medications proposed for use include:  
 
Curcumin - Curcumin is the active ingredient in the Indian spice turmeric and is generally 
recognized as safe (GRAS) by the FDA.  Detailed information about curcumin is found in the 
Research Strategy.  Few side-effects of curcumin have been reported with either 8-12 grams/day 
(8,000-12,000 milligrams/day) of short-term supplementation in adults 61,62,65 and children 64,66,67.  
The primary risk associated with curcumin supplementation is the possibility of temporary/transitory 
gastrointestinal distress (flatulence, constipation, diarrhea, nausea) in a very small percentage of 
individuals and with very high doses 61,65,67.  In the event that symptoms are intolerable, the dose 
will reduced from 25 mg/kg/day to 15 mg/kg/day.  Dr. Chonchol serves as a co-investigator on an 
ongoing NIH-funded trial using curcumin as a treatment of healthy aging (PI: Seals).  As a 
precaution, monthly negative home pregnancy tests will be required in all female participants of 
possible childbearing potential. 
 
Ascorbic Acid (ages 18-25 only) – Similar does of ascorbic acid has been infused intravenously 
without any unfavorable side effects in healthy subjects, as well as patients with chronic disease, 
including cardiovascular disease and chronic kidney disease 68,93,103,104.  However, administration of 
concentrated ascorbic acid may cause irritation to the local area around the infusion.  In order to 

reduce this risk we will dilute the infusion of ascorbic acid in sterile saline.  This procedure is 
currently safely employed in an NIH-funded, IRB-approved protocol in adults with ADPKD.    
 
Nitroglycerin (ages 18-25 only) - Nitroglycerin may cause a slight decrease in arterial blood 
pressure, minor symptoms such as lightheadedness, tingling in the tongue and in the arms and 
legs, headaches, fainting and/or increased heart rate.  The dose of nitroglycerin used is considered 
safe in the proposed population and is also used in patient populations to relieve angina 105.  This 
procedure is currently safely employed in an NIH-funded, IRB-approved protocol in adults with 
ADPKD.  Please see below for monitoring information.    
 
EMLA Cream (as needed) – EMLA cream may cause transient local skin reactions at the 
application site including paleness, erythema, and edema.  Uncommon side effects include a mild 
burning sensation, itching, or warmth at the application side.  In rare cases, allergic reactions or 
anaphylactic shock may occur.   
 
There are no alternative methods that would provide the same type and accuracy of information as the state-
of-the art procedures proposed in this application. 
 
2. Adequacy of Protection Against Risks 
 
a) Recruitment and Informed Consent.  I will use the recruitment and adherence strategies and experience 
previously implemented by myself and my mentorship team.  Dr. Chonchol has successfully recruited patients 
with different kidney diseases, including ADPKD, to clinical trials for many years.  Dr. Cadnapaphornchai has 
been performing research in children and young adults with ADPKD since 2001, including a recently completed 
3 year statin study in 110 children and young adults with ADPKD.  Patients for the study will be recruited from 
the ADPKD research center at the University of Colorado Denver Anschutz Medical Campus (UCD), with 
access to >4800 ADPKD patients around the country, including children and young adults.  Since 1985, UCD 
has maintained an ADPKD registry and mailing list.  Included in this registry are children and young adult 
ADPKD patients from the Denver Metro Area and all over the United States.  In addition to the UCD registry, I 
will advertise through the national PKD Foundation.  The ADPKD center at the UCD has longstanding 
expertise and national recognition, and therefore, is able to attract ADPKD patients from all over the United 
States for clinical studies.  Of note, UCD is currently the only group in the world performing research studies in 
children with ADPKD.  The feasibility of recruitment is supported by the recently completed intervention study 
performed by the co-mentor Dr. Cadnapaphornchai, which included 110 pediatric and young adult patients with 
ADPKD followed for a three-year period, including individuals out of state 56, and these participants will be 
recruited for the proposed research study.  The Division of Renal Diseases and Hypertension at the University 
of Colorado, in general, and Dr. Chonchol and Dr. Cadnapaphornchai specifically, have an excellent track 
record in recruiting participants and meeting enrollment goals for clinical trials.  Dr. Chonchol has served as PI 
or co-PI in many large multi-center clinical trials and will help monitor and guide the progress of the project. 
 
Written informed consent (participants ages 18-25 years of age and parent(s) of children 6-17 years of age) 
and assent (children 6-17 years) will be obtained using a standardized forms approved by the University of 
Colorado Multiple Institutional Review Board (IRB), that provides appropriate information about the study and 
the potential risks and benefits.  The parent(s) and/or child will read the consent/assent form and the 
investigator will review these forms and will answer any questions that the subject may have prior to obtaining 
the subject’s written consent/assent.  The consent form will then be signed by at least one parent and a 
witness for children under 18, or by the participant if 18-25 years of age, as well as by the investigator as 
documentation of consent.  The assent form will also be signed by children 6-17 years of age and by the 
investigator and a witness.  A copy of the signed consent/assent form(s) will be given to the subject/parent(s).  
All of the proposed procedures and protocols will be reviewed and approved by the University of Colorado 
Multiple IRB. 
 
Every attempt will be made to maintain patient’s confidentiality, although a potential risk is loss of 
confidentiality.  Since subjects participating in these studies have already been diagnosed with ADPKD, 
participation is unlikely to affect the ability of to obtain insurance.  The University of Colorado PKD research 
group has 30 years of experience recruiting patients with ADPKD, including patients from out of state.  The 
University of Colorado IRB is aware that our patients travel from out of state and we have taken precautions to 
ensure confidentiality when patients are traveling or getting blood drawn at a contract laboratory (Quest).   
 

b) Protection Against Risk. 
Minimizing General Risks - The potential general risks of the proposed studies will be minimized by: 
•  Screening for adverse events and allergies to drugs. 
• 
Using only safe, well-established procedures, with only qualified and experienced personnel performing the 
procedures. 
• 
Ensuring constant personal monitoring of each experimental session by the investigators and clinical staff. 
• 
Providing appropriate clinical supervision and emergency equipment through the Clinical Vascular 
Physiology Laboratory environment. 
•  Safety monitoring annually by a data safety monitoring board (DSMB). 
• 
Employing record keeping processes with complete confidentiality.  All subject identities and records will 
remain strictly confidential.  Individual subject data will not be associated with subject name. 
• 
Only performing the ascorbic acid infusion and sublingual nitroglycerin administration in a sub-set of young 
adult participants 18-25 years of age. 
• 
All blood pressure, blood draws and FMDBA measurements will be performed in the arm not designated for 
future vascular access for dialysis 
• 
A small venous catheter (for vitamin C intravenous infusion during FMDBA and aortic pulse-wave velocity 
measurements) will be placed in the arm or dorsal aspect of the hand (ages 18-25 only). 
 
Minimizing Specific Risks Related to Drugs/Supplements: 
• 
Risks to the subjects from curcumin administration will be minimized by using a dose and a duration of 
curcumin treatment that have been characterized repeatedly as safe and well tolerated in humans, 
including children 61,62,64,67.  In addition, scheduled safety checks (phone questionnaire) will occur after one 
month, three months, six months, and nine months of taking the curcumin (or placebo).  Serum creatinine, 
ALT and AST will be measured after one month, six months, and nine months, and for those out of state, 
will be performed at a local contracted laboratory (Quest).  Home pregnancy tests will be required monthly 
for all girls of possible childbearing potential (Tanner Stage 2 or higher).  All subjects will be given a digital 
blood pressure monitor (A&D Medical, UA767) with an appropriately sized cuff and the subject/parent will 
be instructed in its use.  Home blood pressures will be taken monthly and reported during phone safety 
checks.  This overall approach has been successfully employed previously by the co-mentor, Dr. 
Cadnapaphornchai 54,56.  In addition, potential side effects will be reviewed with participants at time of 
consent and subjects will be instructed to contact the study coordinator if they experience any side effects 
potentially associated with curcumin therapy. 
• 
Risks of local irritation from the use of ascorbic acid (ages 18-25 only) will be minimized by diluting the 
ascorbic acid in saline.   
• 
Risks associated with sublingual nitroglycerin (ages 18-25 only) will be minimized by screening out 
subjects who take vasodilating drugs that may interact with sublingual nitroglycerin (such subjects will still 
be eligible to participate in all other parts of the study), and not administering nitroglycerin to subjects with 
systolic blood pressure <100-mmHg, mean arterial pressure <70 mmHg, or history of migraines.  During 
nitroglycerin administration, heart rate and blood pressure will be monitored every 2 minutes for signs and 
symptoms of hypotension.  Additionally, an intravenous catheter (also used for ascorbic acid infusion) will 
be in place for administration of normal saline in the rare event of symptoms due to hypotension. 
• 
Risks associated with EMLA cream (as needed) – will be minimized by screening for hypersensitivity to 
prilocaine, lignocaine, or other local anesthetics of the amide type.   
 
Minimizing Specific Risks Related to Confidentiality:  
Strict confidentiality of patients’ information will be kept in accordance of HIPPA policy.  All clinical samples will 
be stored by code alone.  Data will be stored in Research Electronic Data Capture (RedCap), an encrypted, 
secured, HIPPA compliant website that meets all IRB regulations for secure data management.  Paper hard-
copies containing any identifiable information, will be locked in the research offices and in a locked cabinet, 
and will be accessible to members of the research team only on an as-needed basis.  The contract laboratory 
(Quest) will ensure strict confidentiality in accordance of HIPPA for all laboratory data.  All information in our 
database is treated with the same confidentiality as a medical record.         
 
3. Potential Benefits of the Proposed Research to the Subjects and Others 
Because the risks of participating in this study are relatively small, the risk-to-benefit ratio also is relatively low.  
Subjects will receive benefits associated with overall knowledge of their health from any testing performed.  

The results of the study have potential benefits for the treatment of vascular dysfunction in children and young 
adults with ADPKD, and possibly for slowing the progression of renal structural disease, which could potentially 
reduce overall cardiovascular and renal risk and mortality in this population. 
 
4. Importance of the Knowledge to be Gained 
 
The findings from the proposed research should provide important new information regarding the potential 
therapeutic effects of curcumin therapy on endothelial dysfunction and arterial stiffness in children and young 
adults with ADPKD, as there is no available information to date on these questions.  In addition, the proposed 
research should provide important insight into the mechanisms involved in any beneficial effects of curcumin 
on vascular function, as well as exploratory information regarding the influence of curcumin on total kidney 
volume.  Together, this information will contribute to future interventions to treat vascular dysfunction and 
reduce cardiovascular risk in this group of patients. 
 
5. Data Safety Monitoring Plan 
 
a) Medical Supervision and Subject Surveillance.  The trial will be performed by the University of Colorado 
Division of Renal Diseases and Hypertension research group located at the University of Colorado Anschutz 
Medical Campus.  The physicians, advanced practice providers, and nursing staff will have the responsibility of 
performing medical histories and physical examinations as part of the subject screening procedures.  The 
research nursing staff and trained phlebotomists perform the venous catheterizations (ages 18-25 only).  Dr. 
Michel Chonchol, a board certified nephrologist, will provide final decisions regarding subject 
screening/enrollment and will monitor/oversee clinical status and subject safety as they progress through the 
intervention. 
 
b) Data Safety Monitoring Plan.  A data safety monitoring board (DSMB) including clinicians (a nephrologist 
and pediatrician) and a statistician (independent of the study investigators but part of the faculty at the 
University of Colorado, School of Medicine) will be formed to assess potential adverse events.  These data will 
be prepared by the DSMB statistician, ensuring the study statistician remains blinded until the final analysis.  
The DSMB will meet annually to review the protocol and will follow the guidelines established by the NIH 
National Center for Research Resources which include: a) monitoring the progress of the protocol (e.g., 
reviewing subject recruitment, attrition and minority involvement) and the safety of research participants (e.g., 
reviewing unblinded data for safety); b) assuring compliance with requirements regarding the reporting of 
adverse events; c) assuring that any action that results in the temporary or permanent suspension of the 
protocol is reported to all the appropriate monitoring bodies (i.e., IRB, NIH, FDA, etc.); and d) assuring data 
accuracy and protocol compliance. 
 
c) Reporting of Side Effects.  Subjects will be instructed to report side effects to the investigator and their 
nephrologist, if not part of the investigative team.  Any serious adverse events will be reported to the Colorado 
Multiple IRB, Verdure Sciences, and the FDA, in accordance with the guidelines of each of these groups.  
Participants who drop out of the study will be handled using an intent-to-treat analysis. 
 
d) Exit Criteria.  The primary exit criteria will include completion of the study, , patient request, IRB or DSMB 
request, or other rationale as determined by the principal investigator or mentors.  Other exit criteria are listed 
below.  The number of subjects exiting the study and the reasons for exit will be carefully documented. 
 
Patient Stopping Criteria: 
•  Withdrawal of informed consent (subject’s decision to withdraw for any reason) 
•  Any clinical adverse event (AE), laboratory abnormality or intercurrent illness which, in the opinion of the 
investigator, indicates that continued participation in the study is not in the best interest of the subject 
•  Pregnancy 
•  Loss of ability to freely provide consent through imprisonment or involuntary incarceration for treatment of 
either a psychiatric or physical (e.g., infectious disease) illness 
•  Use of an investigational therapy or device other than study medication 
•  Significant non-compliance with protocol (i.e., procedures, assessments, medication, etc.) 
• Serious acute hypersensitivity reactions to investigational drug 
 
Study Stopping Criteria: 
The DSMB may recommend study stopping if: 
 The data show a significantly increased risk of serious adverse effects in the treatment group.   

 It becomes clear that successful completion of the study is not feasible (e.g. there is an excess of patient 
dropout, missing data, lack of recruitment etc). 
 
Clinical Trials.gov Requirements 
The application includes a trial which requires registration at ClinicalTrials.gov.  This registration will be 
performed. 
 
 
 
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