Pilot Study of the Effects of Colchicine in Non-Diabetic Adults With Metabolic Syndrome
Phase: PHASE1
Conditions: Obesity, Metabolic Disease
Sponsor: Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)

1 
 
NICHD Human Subjects Research Protocol 
 
Date of this version: 
August 7, 2018 
Project Number 
14-CH-0119 
 
IND/IDE Number
 
WITHDRAWN120722 
 
IND/IDE Holder  
To: 
Karim Calis, PharmD, Chair, IRB, NICHD  
Recommended by:  
 
, Branch Chief, PEMG, NICHD 
 
 
, Clinical Director, NICHD 
 
 
, Clinical Director, NHLBI 
 
 
, Clinical Director, NIDDK 
 
 
, Clinical Director, CC 
Subject: 
Protocol Amendment: 
Pilot study of the effects of colchicine in non-diabetic adults with 
metabolic syndrome 
 
Identifying words: 
• Disease(s): Metabolic syndrome, obesity, dyslipidemia, inflammation 
• Population:  Non-diabetic adults with Metabolic Syndrome 
• Other:  
colchicine, NLRP3, cryopyrin, NALP3 
 
Principal Investigator: 
*Jack A. Yanovski, MD, PhD 
SGO/DTM/NICHD 
 
Lead Associate Investigator:  *Andrew Demidowich, MD 
OCD/NICHD 
 
Accountable Investigator:  
Jack A. Yanovski, MD, PhD 
SGO/DTM/NICHD 
 
*May obtain consent from participants 

2 
 
Study type (check all that apply): 
                Archived biological specimens/medical information 
                Natural history; definition of phenotype, genotype/phenotype correlation 
       x       Prospective linkage/gene identification, NOT providing information to participants 
                Prospective linkage/gene identification, providing information provided to participants  
                Social science; assessments of knowledge, attitudes and behavior 
                Genetic counseling 
       x       Drugs or devices 
                Gene transfer 
                Other interventions 
 
  
Estimated Duration of study: 3 years 
 
Subjects of Study: 
   
 
 
 
  
  
Description 
Number 
Sex 
Age 
Adults screened for protocol 
- 
Screened but not enrolled 
- 
Non-diabetic obese adults with metabolic 
syndrome randomized to colchicine or placebo 
- 
Studied, but not randomized (Evaluation-only) 
- 
Open-Label T2DM arm 
250 
163 
40 
 
40 
7 
M, F 
 ≥ 18 
 
Project uses ionizing radiation: Yes 
 
Medically indicated or research? Research 
 
Project involves use of durable power of attorney: No 
 
Offsite project: No 
 
  
 
 
Research participants to be seen at: 
      x        NIH only*  
 
 
                Off-site only 
 
 
                Both NIH* and off-site 
*Includes participants who physically come to the NIH Clinical Center and/or for whom 
specimens/data are analyzed by Clinical Center departments. If participants will be seen 
at NIH, a Medical Advisory Investigator must be indicated on the 1195 form, unless this 
is a social science project with no clinical interventions. 
 
 
Collaborative Project: 
No. 
 
 

3 
 
Précis: 
 
Obesity affects one-third of the adult U.S. population and is a major risk factor for the development 
of type 2 diabetes and cardiovascular disease.  Mouse models and human data suggest that obesity-induced 
chronic inflammation is one mechanism promoting obesity-associated comorbid conditions.  In obesity, 
innate immunity is activated by circulating molecules such as fatty acids and cholesterol crystals bind to 
nucleotide-binding oligomerization (NOD)-like receptor family, pyrin domain containing 3 (NLRP3) 
receptors in adipocyte tissue macrophages (ATMs).  This binding stimulates NLRP3 oligomerization, 
inflammasome formation, and proinflammatory cytokine activation.  The resultant inflammatory cascade 
leads to insulin resistance and decreased pancreatic beta-cell reserve.  It has been proposed that the 
suppression of this chronic low-level inflammatory state may impede the onset of diabetes and 
cardiovascular disease. 
Recent studies have shown colchicine, a potent microtubule inhibitor commonly used for the 
treatment of gout and some rare inflammatory conditions, disrupts intracellular localization of NLRP3, 
thereby blocking inflammasome assembly.  As there are limited medical therapies proven effective to 
improve obesity-related metabolic dysregulation, we propose to determine the efficacy of colchicine 0.6 mg 
twice daily in non-diabetic obese adults with metabolic syndrome. We will conduct a randomized, double-
blinded, placebo-controlled pilot trial of colchicine in forty subjects.  We will study changes in insulin 
resistance, beta-cell reserve, and systemic inflammation.  Using adipose tissue obtained from biopsies, we 
will also study colchicine’s local effects on inflammation and insulin resistance.  Should results prove 
promising, this pilot study will allow determination of the sample size needed for an adequately powered 
study of the effects of colchicine in obese adults with metabolic syndrome. 
Seven patients with type 2 diabetes will be given open-label colchicine and followed as described 
above. We also plan to perform baseline evaluations on 40 subjects who are not eligible for the treatment 
protocol. This group will consist of non-obese adults, obese adults who are not insulin-resistant, and adults 
with diet-controlled type 2 diabetes.  

4 
 
Table of Contents 
 
I. 
Introduction 
6 
II. 
Study Objectives 
13 
III. 
Specific Aims and Hypotheses 
14 
IV. 
Study Design and Methods 
15 
A. Subject Eligibility 
15 
B. Subject Enrollment 
20 
C. and D. Clinical evaluation and Research Testing/Treatment 
20 
E. Biological Specimens 
26 
F. Molecular Genetic Analysis of DNA 
29 
G. Approved Drugs 
29 
H. Unapproved Drugs 
29 
I. Sharing of Data with Subjects 
29 
J. Questionnaires/Psychological instruments 
30 
K. Genetic Counseling 
30 
L. Follow-up of Patients 
30 
M. Conditions for Discharge from Protocol 
30 
V. 
Analysis of the Study/Oversight 
31 
A. Clinical Description 
31 
i.  
Study Administration, Data Collection, Management, and Storage 
31 
ii.  
Publication of Research Findings 
32 
iii.  
Statistical Analysis of the Data 
32 
iv.  
Monitoring Subjects and Criteria for Withdrawal 
33 
v. 
Stopping Rules 
33 
B. Molecular Studies 
34 

5 
 
VI. 
Determination of Sample Size 
34 
VII. 
Human Subjects Protection 
35 
A. Rationale for Subject Selection 
35 
B. Participation of Vulnerable Populations 
36 
C. Screening Methodology 
36 
D. Risk/Benefit Analysis 
37 
E. Privacy and Confidentiality Provisions 
45 
F. Compensation 
46 
G. Consent Procedures 
46 
H. Pharmaceutical, Biologic, and Device Info 
47 
I. Recruitment Strategies 
49 
J. Existing Data 
49 
K. Criteria for Withdrawal from Study 
49 
L. Re-evaluation of Study 
51 
M. Collection, monitoring, analysis and reporting of adverse events 
51 
a. Waiver of Reporting of Anticipated Non-UP adverse events and 
minor protocol deviations 
52 
b. Plan to monitor and report adverse events 
54 
c. DSMB 
54 
N. IND Monitoring Plan 
54 
VIII. 
Appendices 
58 
IX. 
References 
88 

6 
 
Introduction: 
 
Obesity is a serious disorder that affects approximately one-third of the adult U.S. population and is 
a major risk factor in the development of type 2 diabetes (T2DM), hypertension, dyslipidemia, and 
cardiovascular disease.1-3 The pathophysiology underlying the relationship between obesity, insulin 
resistance, and cardiovascular disease is not fully understood, but numerous studies give evidence that 
inflammation plays a prominent role.  
 
Mouse models of diet-induced and genetically-induced obesity show increased levels of 
inflammatory cytokines, including interleukin (IL)-1β, and IL-18.4  Further, injecting wild-type mice with 
IL-1β decreases insulin sensitivity, whereas IL-1β null mice are protected from insulin resistance after a 12 
week high fat diet (HFD)5.   
Cross-sectional human studies also implicate inflammation in the development of insulin resistance 
and cardiovascular disease.  Data from NHANES III indicate that inflammation is positively correlated with 
BMI6 and the presence of metabolic syndrome (MetS).  Individuals with MetS had more inflammation than 
those without MetS. Higher C-reactive protein (CRP) levels were seen with increasing components of MetS.  
Inflammatory cytokine levels, such as IL-1β, IL-6, and IL-18, were also positively correlated with BMI.7  In 
the Women’s Health Initiative trial, elevated levels of hsCRP and IL-6 were associated with increased risk 
for T2DM and vascular events in apparently healthy postmenopausal women.8,9  Conversely a reduction in 
body weight, either via lifestyle modification or bariatric surgery, lead to decreased levels of inflammation 
and insulin resistance.10,11  Other large cross-sectional studies have also confirmed a relationship between 
inflammation and metabolic abnormalities12,13 and the development of coronary artery disease.14-17 
 
The sources of the chronic low-grade inflammation of obesity arise from multiple organs, but it 
appears that adipose tissue (AT) itself is a large contributor to the inflammatory process.  AT consists of 
adipocytes and the stromal vascular fraction (SVF), which includes fibroblasts, vascular tissue, and adipose 
tissue macrophages (ATM).  In lean individuals, macrophages comprise less than 10% of the cells in AT.18  
The fat cells themselves are smaller in volume and secrete higher levels of adiponectin and omega-3 fatty 
acids, which predispose the macrophages to differentiate to an anti-inflammatory M2 state.  M2 macrophages 
release immunosuppressive cytokines, such as IL-10 and TGFβ, which act on fat and muscle tissue to 
enhance insulin sensitivity.19 However, in obese individuals, adipocytes are both greater in volume and 
number, leading to inefficient triglyceride storage and increased levels of circulating fatty acids and 
ceramides.20  These molecules attach to the pattern recognition receptors in ATMs, which stimulate 
differentiation to the pro-inflammatory M1 phenotype.21  Chronic inflammation ensues, with an increased 
release of chemokines and cytokines including TNFα, IL-1β, IL-6, IL-18, plasminogen activator inhibitor-1 
(PAI-1), monocyte chemoattractant protein-1 (MCP-1), and reactive oxygen species (ROS).21-23  These 

7 
 
factors further augment adipose tissue inflammation by recruiting additional ATMs as well as B and T 
cells.24    
The resultant inflammation is thought to lead to insulin resistance through several concomitant 
pathways.  GLUT4 mediates glucose uptake in adipocytes and skeletal muscle and is integral in glucose 
homeostasis.  In insulin resistant (IR) states, GLUT4 expression in AT is downregulated, and insulin-
stimulated GLUT4 transport to the plasma membrane is impaired.25  Additionally, the chronic inflammatory 
state leads to serine phosphorylation and deactivation of insulin receptor substrate-1 (IRS-1), an important 
downstream mediator of the insulin receptor, by IKK-β.26  IRS-1 expression is downregulated as well.   
Interestingly, Lumeng et al observed that in co-cultured adipocyte and macrophage cells this phenomenon 
was partially reversed when treated with anti-inflammatory TNFα neutralizing antibodies.25    
Moreover, low-grade systemic inflammation can impair pancreatic islet cell functioning and insulin 
secretion.  In early stages of β-cell dysfunction, inflammatory cytokines disrupt the proper intracellular 
calcium storage and flux necessary for adequate insulin secretion.27,28  Chronically, inflammatory signaling 
via the NF-kB and mitogen-activated protein kinase (MAPK) pathways leads to mitochondrial stress, 
reactive oxygen species (ROS) formation, and eventual β-cell apoptosis.29,30 
 
NLRP3 mediates inflammatory activation in obesity 
 
Leukocytes express pattern recognition receptors on their cell surface which allow for induction of a 
rapid inflammatory response to pathogen-associated molecular patterns.  These components of the innate 
immune system, allow for rapid activation of cell-signaling pathways and production of a pro-inflammatory 
cytokine cascade in response to infection.  Similarly, members of the NOD-like receptor (NLR) family can 
recognize endogenous molecules (e.g. free fatty acids, ceramides, urate crystals, ATP) released during cell 
damage and stimulate a robust inflammatory response.31 
 
A member of the NLR family, the NLRP3 (Nod-like receptor family, pyrin domain containing 3) 
inflammasome has been implicated as the major source of chronic inflammation seen in obesity.  NLRP3 
(also known as cryopyrin or NALP3) is primarily expressed in monocytes and macrophages, with little 
production seen in adipocytes or other leukocytes.11,32  Upon binding with a “danger signal,” NLRP3 
receptors oligomerize in the cytosol with procaspase-1 via the adaptor protein ASC (apoptosis-associated 
speck-like protein containing a caspase recruitment domain; also known as PYCARD).  The formation of 
this inflammasome autocatalytically cleaves the cysteine protease procaspase-1 to its active form, caspase-1, 
which in turn cleaves inactive pro-IL-1 and pro-IL-18 into their active forms.33  IL-1β initiates the acute 
phase response and triggers the production and secretion of other pro-inflammatory cytokines, such as TNFα, 

8 
 
IL-6, and MCP-1, resulting in a progressively amplified cytokine network.34  IL-18 also induces the 
production of a number of cytokines, chemokines, and cell-adhesion molecules.35 
 
Obesity and T2DM are associated with elevated circulating levels of danger-associated molecular 
patterns (DAMPs), such as free fatty acids and ceramides.36,37  When human macrophages are cultured with 
saturated fatty acids, IL-1β and IL-18 production increases, but such increases can be abrogated in 
macrophages knocked out for NLRP3, ASC, or caspase-1.  However, knocking out a different Nod-like 
receptor, NLRC4, has no effect.5  Likewise, ceramides, derived from the metabolism of long-chain saturated 
fatty acids, also activate caspase-1 and IL-1β production in human wild-type macrophages, but not in 
NLRP3-knockouts.11 
In vivo mouse models demonstrated similar results.  Nlrp3 and IL1b expression in AT is positively 
correlated with body weight and adiposity in mice, and food restriction results in significant decreases in 
their expression.11  Knocking out components of the NLRP3 inflammasome (e.g. Nlrp3-/-, ASC-/-, caspase-1-/- 
and lL1b-/- mice) does not affect baseline glucose metabolism and insulin sensitivity, but prevents the 
development of insulin resistance and inflammatory cytokine expression during a protracted HFD state.4,5,11,38  
Additionally, ablation of the NLPR3 inflammasome results in markedly smaller adipocytes, elevated levels 
of adipose GLUT4 and adiponectin, with decreased inhibition of IRS-1.11,39  Caspase-1 knockouts also 
demonstrate increased fat oxidation and gain significantly less bodyweight than wild type mice during a 10-
week HFD regimen, despite similar food intake.4  Interestingly no changes in triglyceride or cholesterol 
levels are seen in obese Nlrp3 knockout mice.11 
In humans, metabolically unhealthy obese individuals have greater levels of NLRP3 activity, IL-1β 
expression, AT macrophages, and regulatory T cells (Tregs) in their visceral adipose tissue (VAT) than either 
metabolically healthy obese or lean individuals.   Weight loss in obese subjects with T2DM leads to 
enhanced insulin sensitivity and decreased mRNA expression of IL1B and NLRP3 in subcutaneous adipose 
tissue (SAT).11 
Other circulating molecules seen in models of diet-induced obesity have been found to be potent 
NLRP3 activators.  Oxidized low-density lipoprotein (LDL) and cholesterol crystals increase NLRP3 
inflammasome formation in mice, leading to macrophage activation and IL-1β secretion.  Conversely, 
ablation of the inflammasome reduces early atherosclerosis in high cholesterol diet-fed mice.40  Monosodium 
urate (MSU) crystals, seen in gout and obesity, can also act as a danger signal by stimulating the NLRP3 
inflammasome.41,42   
The progression from insulin resistance to T2DM involves pancreatic inflammation, destruction, and 
reduced pancreatic β-cell reserve.43,44  In a study by Youm, et al, chronically DIO mice had greater levels of 
inflammation, islet fibrosis, and β-cell death than their lean chow-fed littermates, whereas DIO Nlrp3-/- mice 
were protected from these untoward effects.  Consistent with this finding, the DIO Nlrp3-/-mice had lower 

9 
 
fasting glucose and higher fasting insulin levels than their WT counterparts, suggesting that NLRP3 plays an 
important role in both early (i.e. peripheral insulin resistance) and late (i.e. pancreatic beta cell destruction) 
stages of metabolic dysfunction.38 
 
Anti-inflammatory agents in obesity 
 
 
If inflammation is the root cause of obesity-related metabolic dysregulation, then anti-inflammatory 
agents should show benefit.  This has been the case for some mouse models of obesity.  HFD mice treated 
with a neutralizing IL-1β antibody demonstrate significant improvement of glycemic control and beta cell 
function,29 with no effect on weight gain or food intake.45  Given prophylactically, IL-1β antibody treatment 
helps prevent the onset of fasting hyperglycemia and insulin resistance in HFD mice.  IL-1 receptor 
antagonist (IL-1Ra) treatment of DIO mice leads to similar results, with decreased β-cell apoptosis, increased 
β-cell proliferation, and improved glucose-stimulated insulin secretion.45,46  Studies inducing blockade of 
other inflammatory cytokines or pathways in mice, including TNFα47, IL-648, MCP-149, and NF-kB50 have 
also demonstrated favorable metabolic results from reduction of inflammatory processes. 
 
However, such robust results have not been replicated in human clinical trials.  Blockade of TNFα 
had no effect on metabolic parameters T2DM participants47 and only marginal impact in non-diabetic insulin 
resistant subjects.51  Similar findings were seen when using aspirin or the IL-1β antibody canakinumab.52-54  
A different monoclonal IL-1β antibody, gevokizumab, has led to improvements in HbA1c, but not beta-cell 
secretory function or insulin sensitivity in preliminary trials.55  Only anakinra, a recombinant human IL-1Ra, 
has demonstrated any significant benefit, with an improvement in beta-cell secretory function and decrease in 
Hemoglobin A1c in diabetic subjects,56 but not in subjects with impaired glucose tolerance.57 Furthermore, 
anakinra did not affect insulin sensitivity as measured by insulin clamp or HOMA-IR in either study.  
 
While these results are disappointing, they are not wholly unexpected. The machinery involved in 
obesity-induced chronic inflammation is complex.  Cytokines may play complementary, if not redundant, 
roles in promoting inflammation.  Chemokines and endothelial adhesion molecules play integral roles in the 
sustained inflammatory response by facilitating chemoattraction, diapedesis, and migration of additional 
leukocytes to adipose and pancreatic tissues.  For this reason, eliminating a single cytokine may not be 
sufficient to demonstrate significant changes in glucose homeostasis. 
 
Colchicine in preclinical trials 
 
Colchicine remains an intriguing treatment for obesity-associated chronic inflammation because it 
mitigates inflammation through several mechanisms.  Colchicine is a tricyclic alkaloid, extracted from the 

10 
 
flowering plants Colchicum autumnale and Gloriosa superba.58  It has been used to treat gout for over 2000 
years, although the purified form was brought to prominence at the end of the 19th century.59  Colchicine 
exerts its effects by binding in a poorly reversible manner to tubulin, thereby interfering with microtubule 
formation and elongation.  At higher concentrations colchicine promotes depolymerization of microtubules.60  
 
Classically, colchicine was thought to exert its anti-inflammatory actions by inhibiting neutrophil 
chemotaxis and diminishing release of lysosomal enzymes during phagocytosis.  Colchicine also decreases 
leukocyte recruitment by blocking the production and/or release of chemotactic molecules leukotriene B4 
and crystal-derived chemotactic factor (CCF).61   Leukocyte recruitment is further impaired by the inhibition 
of adhesion molecules, such as E-selectin and P-selectins on endothelial cells, L-selectin on leukocytes, 
vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1)61,62.  Surface 
expression of the TNFα receptor in macrophages is also reduced with colchicine, as is neutrophil superoxide 
anion production.63,64  Recent studies also suggest that colchicine may modulate expression of genes involved 
in inflammation or cell migration.65,66  
Notably, oligomerization and activation of the NLRP3 inflammasome is disrupted by 
colchicine.  Microtubules mediate NLRP3 inflammasome formation by transporting ASC and NLRP3 into 
close proximity to each other in the cell, and inhibition of microtubule formation by colchicine has been 
shown to significantly inhibit NLRP3 activation in macrophages both in vitro and in vivo.41,67 
 
Colchicine in human studies 
 
 
Colchicine is currently FDA-approved for the treatment of gout and Familial Mediterranean Fever,68 
and has shown efficacy in other inflammatory conditions.69-71 As inflammation has been hypothesized to play 
a role in the development of cardiovascular disease, several clinical studies have recently examined the 
efficacy of colchicine in cardiovascular-related conditions.  Colchicine 0.5 mg twice daily decreased hsCRP 
levels after one month of treatment in subjects with stable coronary artery disease already on a statin and 
aspirin,72 although this was not seen in patients with acute coronary syndrome or acute stroke.73  Subjects 
with paroxysmal atrial fibrillation, randomized to colchicine after radiofrequency ablation, had decreased 
atrial fibrillation recurrence at 3 months and more substantial post-treatment decrements in hsCRP and IL-6 
levels than those randomized to placebo.74 Colchicine has also demonstrated considerable benefit in reducing 
incidence of recurrent pericarditis75-77 and post-pericardectomy syndrome.78,79 
 
Notably, colchicine has great promise preventing acute coronary syndrome in high risk individuals.  
Among the 532 subjects with stable coronary artery disease on a statin and aspirin and/or clopidogrel, 30% 
of whom had Type 2 diabetes mellitus (T2DM), those randomized to colchicine demonstrated a significantly 
decreased primary composite endpoint of acute coronary syndrome, out-of-hospital cardiac arrest or stroke at 

11 
 
3 years as compared to placebo.80  Another study examined restenosis rate in patients with T2DM 
undergoing percutaneous coronary intervention with bare metal stent placement.  Those randomized to 
colchicine had significantly fewer restenotic events and less neointimal hyperplasia as compared to subjects 
taking placebo.81  Subjects in these studies, many of whom had features of the metabolic syndrome, tolerated 
colchicine well, with the most common side effect being diarrhea/gastrointestinal disturbances (7 – 16%).  
In addition to predisposing to cardiovascular disease, inflammation is thought to play a role in 
insulin resistance.  However, colchicine’s effect on glucose or insulin in individuals with metabolic 
dysregulation is unknown.  Early studies on colchicine suggested that it worsened glucose control by 
inhibiting early and late phase insulin secretion.  However, these studies were done in healthy adults, and the 
treatment duration was less than two weeks.82,83  In subjects with Familial Mediterranean Fever chronically 
on colchicine for 2-13 years, data from frequently-sampled intravenous glucose tolerance testing (FSIVGTT) 
and oral glucose tolerance testing did not demonstrate any differences in fasting insulin or glucose levels, 
early-phase insulin release, or glucose dynamics, as compared with age-matched healthy volunteers.84  
Additionally, neither a Cochrane Database review nor the Colcrys prescriber information insert show 
hyperglycemia as a potential adverse effect of colchicine use.68,85 
 
By blocking NLRP3 inflammasome formation and cytokine production, a plausible mechanism 
exists by which colchicine could improve glucose homeostasis and insulin resistance in obese individuals 
who have evidence for inflammation.  Improvement in metabolic dysregulation may also improve 
cardiovascular risk factors in individuals with MetS, although this has not yet been formally evaluated using 
colchicine.  Consequently, colchicine is an intriguing treatment option for metabolic dysregulation and 
warrants investigation to determine its potential health benefits in obese insulin-resistant individuals. The 
results of such a study also have the potential to advance our knowledge regarding the interplay of 
inflammation and metabolic dysregulation in man.  
 
Adipocyte Ex-Vivo Studies 
 
Whole body insulin-mediated glucose uptake depends primarily on glucose uptake in three organ 
systems: liver, adipose, and muscle.  Upon binding to the insulin receptors in peripheral tissues, insulin 
signaling increases the number of insulin-stimulated glucose transporters (GLUT4) at the cell surface and 
allows augmented glucose translocation across adipocyte and muscle plasma membranes.  However, most 
research on the kinetics of insulin signaling pathways and the studies that examine how inflammation 
perturbs these pathways have been performed largely in either non-human models86-88 or cultured 3T3-L1 
mouse cells.89-91 Preliminary studies have shown that these kinetic models are inadequate to capture insulin 
dynamics at the level of the human adipocyte because they do not recapitulate the multiple phases of GLUT4 
trafficking. While in vivo human studies would serve as the most appropriate representation of insulin 

12 
 
kinetics, the logistics of these measurements make them unrealistic. Therefore, ex vivo experiments may 
instead serve as a more reasonable portrayal of the kinetics of the adipocyte’s response to insulin within 
obese insulin-resistant subjects with chronic inflammation. 
 
Thus, collaborating with the NICHD Section on Membrane and Cellular Biophysics as a part of the 
NICHD Director’s Investigator Award, we have initiated investigations of the effects of colchicine on insulin 
sensitivity locally in the subcutaneous adipose tissue. Dr. Chad McCormick, a member of this collaboration, 
is examining insulin kinetics in coded adipocyte biopsy samples obtained from our subjects by creating a 
novel reporter system in which we transfect subjects’ adipocytes with a HA-pHTomato-GLUT4-mEGFP 
plasmid to quantify GLUT4 translocation to the cell surface after maximal insulin stimulation in culture.  
 
Additionally, it has been recently hypothesized that individual adipocytes do not exhibit variable 
degrees of insulin sensitivity along a spectrum, but rather manifest a binary insulin-sensitive or insulin-
resistant state.92 Consequently, insulin-resistant subjects have some insulin-sensitive adipocytes that respond 
to insulin in a similar fashion as those of lean, healthy individuals, but the percentage of responsive cells in 
insulin-resistant individuals is much lower than that of lean, healthy individuals.92 Dr. McCormick is 
utilizing quantitative immunofluorescence microscopy to investigate whether local SAT inflammation 
promotes IR states by changing insulin-sensitive adipocytes into insulin-refractory adipocytes [Figure 1]. As 
Akt is phosphorylated upon insulin binding to the insulin receptor, we are able to visualize and quantify 
phospho-Akt (phosphorylation at Thr308; pThr308). Ultimately this allows us to evaluate insulin sensitivity 
at the cellular level. By also staining the tissue with anti-CD11c antibody to identify M1 pro-inflammatory 
macrophages as well as CD68, a universal macrophage marker, we are able to assess whether adipocytes in 
close proximity to activated macrophages lack phospho-Akt (pThr308) and therefore exhibit suppressed 
insulin signaling.  
 
 

13 
 
  
 
 
 
The subjects evaluated by the aforementioned methods in our protocol thus far consist only of obese 
insulin-resistant non-diabetics.  Because of the potentially significant findings and novel strategies to 
evaluate local insulin sensitivity in human SAT, it would be worthwhile to investigate how these results 
compare with healthy non-insulin resistant volunteers and subjects with T2DM.   
 
II. Study Objectives: 
 
We propose a pilot study of the effects of colchicine in obese non-diabetic adults with metabolic 
syndrome to determine if colchicine treatment can improve metabolic and inflammatory parameters, both 
locally and systemically, independent of weight loss. This pilot study will determine the sample size needed 
for an adequately powered larger study in the future. 
 
We also propose enrolling additional subjects who do not qualify for the protocol to evaluation only. 
This group will consist of non-obese and obese adults without significant insulin resistance and/or 
inflammation as well as subjects with diet-controlled T2DM. They will not be randomized or receive study 
medication.  The results from these controls will allow us to determine insulin kinetics in SAT over the entire 
spectrum of insulin sensitivity.  
 
Subjects with T2DM have more severe metabolic abnormalities and typically higher inflammatory 
levels, but their pancreatic function may possibly be past the point of reversibility.  Therefore, it is unclear 
whether such subjects would be the most likely or least likely group to derive benefit, if any, from colchicine.  
We propose to enroll seven diet-controlled subjects with T2DM to an open-label colchicine arm. Results 
A 
B 
Figure 1. (A) Immunofluorescence imaging of a 3µm Z-slice of subcutaneous adipose tissue from a subject. Blue 
staining represents AKT p308 (insulin sensitivity), red represents CD68 (macrophages), and green represents 
CD11c (M1 pro-inflammatory macrophages). Most adipocytes have Akt p308 staining, but some are notably 
absent. (B) Zoomed-in images of activated M1 macrophages on Bright field and confocal merge 
immunofluorescent microscopy.  Many of the adipocytes surrounding the M1 macrophages lack Akt p308 staining 
and are insulin-refractory. 

14 
 
from this arm will allow us to determine the sample size that might be needed if a future study for the effects 
of colchicine in T2DM were to be carried out. 
 
III. Specific Aims and Hypotheses:  
 
Through this randomized, double-blind, placebo-controlled clinical trial, we will obtain pilot data for the 
following aims and hypotheses: 
 
1. 
Aim 1: To explore changes in insulin sensitivity, as measured by FSIVGTT, among obese 
non-diabetic adults following long-term colchicine administration. 
Hypothesis 1: We hypothesize that obese non-diabetic adults on colchicine 0.6mg bid for 3 
months will exhibit significant improvements in insulin sensitivity compared with placebo, 
independent of weight loss.  We also hypothesize that reductions in systemic inflammation, 
as measured by hsCRP, will moderate this relationship. 
2. 
Aim 2: To explore changes in other parameters of glucose homeostasis, among obese non-
diabetic adults following long-term colchicine administration. 
Hypothesis 2: Using data from the FSIVGTT, we hypothesize that obese non-diabetic adults 
on colchicine will exhibit significant improvements in acute pancreatic reserve (as measured 
by acute insulin response to glucose (AIRg)) and glucose homeostasis (as measured by the 
Disposition Index). We also hypothesize that insulin resistance as measured by HOMA-IR 
will demonstrate improvement in colchicine-treated subjects.  
3. 
Aim 3: To explore changes in cardiovascular risk factors among obese non-diabetic adults 
following long-term colchicine administration. 
Hypothesis 3: We hypothesize that colchicine-treated subjects will demonstrate significant 
improvements in cardiovascular risk factors and other lipid parameters, including blood 
pressure, LDL, HDL, Lp(a), fatty acid levels, uric acid, hemoglobin A1c, and fasting plasma 
glucose as compared with placebo-treated subjects. 
4. 
Aim 4: To explore changes in inflammation among obese non-diabetic adults following 
long-term colchicine administration. 
Hypothesis 4a: We hypothesize that colchicine-treated subjects will demonstrate significant 
improvements in systemic inflammatory markers as well as leukocyte/endothelial adhesion 
markers. 
Hypothesis 4b: We hypothesize that colchicine-treated subjects will demonstrate significant 
improvements in local adipose tissue inflammation. 

15 
 
5. 
Aim 5: To explore insulin-dependent GLUT4 dynamics locally in human adipocytes across 
the spectrum of insulin sensitivity.  
Hypothesis 5a: We hypothesize that colchicine-treated subjects will demonstrate significant 
improvements in adipose insulin-dependent GLUT4 kinetics in vitro. 
 
IV. Study Design and Methods: 
 
A. Subject Eligibility: All adults who meet the inclusion and exclusion criteria as listed below and who 
can attend all needed study visits at the NIH Clinical Center will be eligible: 
 
I. 
Inclusion Criteria for subjects randomized to colchicine or placebo: 
 
Subjects will qualify for randomization to colchicine or placebo if they meet the following criteria: 
1. 
Good general health.  In general subjects should take no medications.  However, individuals 
taking medications for obesity-related co-morbid conditions, who have not had changes in 
dosage for more than 3 months, may be included, at the discretion of the principal 
investigator. 
2. 
Obesity, defined as a body mass index (BMI) ≥ 30 kg/m2, but weight less than 450 lbs in 
order for subjects to undergo Dual-Energy X-ray Absorptiometry (DXA) scanning. 
3.  
Age 18-100 years. 
4.  
Metabolic Syndrome93 
(Any 3 of the following 5): 
a. 
FPG ≥ 100 mg/dl or Impaired Glucose Tolerance (Glucose ≥ 140 mg/dl at 2 hours of 
OGTT) 
b. 
Triglycerides ≥ 150 mg/dl, or on treatment 
c. 
Waist Circumference: Men ≥ 40 in (≥ 102 cm); Women ≥ 35 in (≥ 88 cm) 
d. 
Reduced HDL-C: Men < 40 mg/dl; Women < 50 mg/dl, or on treatment 
e. 
Hypertension: ≥130 mmHg systolic, or ≥85 mmHg diastolic, or on treatment 
5. 
HOMA-IR ≥ 2.6.  Our goal is to enroll participants who have pre-existing insulin 
resistance.94 
6. 
hsCRP ≥ 2.0 mg/L. We aim to recruit participants with increased baseline level of 
inflammation. Individuals with hsCRP above 2.0 mg/L have been shown to have an 
increased risk for cardiovascular events.16  
 
II. 
Exclusion Criteria for subjects randomized to colchicine or placebo:  

16 
 
1. 
Type 2 diabetes mellitus, as determined by either having: 
a. 
“clear clinical diagnosis” of diabetes, such as “a patient in a hyperglycemic crisis or 
classic symptoms of hyperglycemia and a random plasma glucose ≥200 mg/dL” 
b. 
two of the following three: 
 
i. 
fasting plasma glucose ≥ 126 mg/dL 
ii. 
Hemoglobin A1c ≥ 6.5% 
iii. 
An oral glucose tolerance test glucose concentration of ≥ 200 mg/dL at 2 
hours. 
c. 
one of the above three criteria (bi.-biii.) meeting the T2DM cutoff on two different 
days. 
If only one of the above three criteria (bi.-biii.) meet the T2DM threshold during the 
Screening Visit, that test will be repeated on another day to determine if the subject has 
T2DM or not.  As per ADA guidelines, “The diagnosis [of T2DM] is made on the basis of 
the confirmed test.”95 
 
Moreover, because HbA1c has been shown to be higher in African Americans (AA) 
as compared to other races for the same glycemia, non-diabetic AA may be unfairly 
excluded by their HbA1c alone.96-98  Therefore, for AA subjects, if their 2 hour OGTT and 
fasting glucoses are in the non-diabetic range, and the HbA1c is < 7.0%, we will consider 
them non-diabetic. 
2. 
Presence of a significant active or chronic illness likely to limit life span and/or increase risk 
of intervention, including renal (GFR ≤ 60 ml/min/1.73m2), cardiovascular, hepatic (other 
than obesity-related steatosis), gastrointestinal, immunologic, endocrinologic (e.g. Cushing 
syndrome), pulmonary (other than either asthma not requiring continuous medication or 
sleep apnea-related disorders), or other disorders at the discretion of the investigators. 
3. 
Recent use of colchicine or anorexiant medications in the last 3 months. 
4. 
Known allergy to colchicine. 
5. 
Previous history of agranulocytosis, gout, or significant myositis. 
6. 
Females who are pregnant, planning to become pregnant, currently nursing an infant, or have 
irregular menses, defined as cycles less than 21 days or greater than 45 days.99 
7. 
Individuals who have current substance abuse or a psychiatric disorder or any other 
condition that in the opinion of the investigators would impede competence, compliance, or 
participation in the study. 
8. 
Subjects who regularly use prescription medications unrelated to the complications of 
obesity, especially those known to affect enzymes involved in colchicine metabolism, such 

17 
 
as CYP3A4 or P-glycoprotein (P-gp) (see Appendix B1).  Oral contraceptive use will be 
permitted, provided the contraceptive has been used for at least two months before starting 
study medication.  The use of over-the-counter and prescription medications will be 
reviewed on a case-by-case basis; depending on the medication, subjects who have 
continued to take prescription medication or have stopped taking an exclusionary medication 
for at least 3 months prior to study entry may be eligible (see Appendix B2). 
9. 
Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight 
change of more than 3% of body weight in the past two months. 
10. 
Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the last 
7 days prior to fat biopsy.  
11. 
History of keloid formation. 
12. 
Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes). 
 
III. 
Inclusion Criteria for subjects who are evaluated but not eligible for randomization: 
 
Subjects will qualify for the Evaluation-only arm if they meet the following criteria: 
1. 
Good general health.  In general subjects should take no medications.  The use of over-the-
counter and prescription medications will be reviewed on a case-by-case basis; depending on 
the medication, subjects who have continued to take prescription medication or have stopped 
taking an exclusionary medication for at least 3 months prior to study entry may be eligible 
(see Appendix B2). 
2. 
Weight less than 450 lbs in order for subjects to undergo Dual-Energy X-ray Absorptiometry 
(DXA) scanning. 
3.  
Age 18-100 years. 
 
IV. 
Exclusion Criteria for subjects who are evaluated but not eligible for randomization:  
1. 
Type 2 diabetes mellitus that is not well controlled with diet alone: subjects taking an anti-
diabetic medication (e.g. metformin, insulin, sulfonylureas, etc.) or having a Hemoglobin 
A1c > 9.0% 
2. 
Presence of a significant active or chronic illness likely to limit life span and/or increase risk 
of intervention, including renal (GFR ≤ 60 ml/min/1.73m2), cardiovascular, hepatic (other 
than obesity-related steatosis), gastrointestinal, immunologic, endocrinologic (e.g. Cushing 
syndrome), pulmonary (other than either asthma not requiring continuous medication or 
sleep apnea-related disorders), or other disorders at the discretion of the investigators. 
3. 
Recent use of colchicine or anorexiant medications in the last 3 months. 

18 
 
4. 
Females who are pregnant, planning to become pregnant, or are currently nursing an infant.  
5. 
Individuals who have current substance abuse or a psychiatric disorder or any other 
condition that in the opinion of the investigators would impede competence, compliance, or 
participation in the study. 
6. 
Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight 
change of more than 3% of body weight in the past two months. 
7. 
Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the last 
7 days prior to fat biopsy.  
8. 
History of keloid formation. 
9. 
Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes). 
 
V. 
Inclusion Criteria for subjects with diet-controlled T2DM 
Subjects will qualify for the Open Label arm if they meet the following criteria  
1. 
Diet-controlled T2DM, as determined by having all of the following: 
a  
A diagnosis of T2DM (as defined above in Section IV.II.1.). 
b  
Not on any diabetic/hypoglycemic agents 
c  
Not having an alternate cause of hyperglycemia (e.g. T1DM, glucocorticoid-
induced, lipodystrophy, acromegaly, etc.) 
d 
Hemoglobin A1c ≤ 9.0% 
2. 
Good general health.  In general subjects should take no medications.  The use of over-the-
counter and prescription medications will be reviewed on a case-by-case basis; depending on 
the medication, subjects who have continued to take prescription medication or have stopped 
taking an exclusionary medication for at least 3 months prior to study entry may be eligible 
(see Appendix B2). 
3.  
Age 18-100 years. 
4. 
Obesity, defined as a body mass index (BMI) ≥ 30 kg/m2, but weight less than 450 lbs in 
order for subjects to undergo Dual-Energy X-ray Absorptiometry (DXA) scanning. 
5.  
Metabolic Syndrome93 
(Any 3 of the following 5): 
a. 
FPG ≥ 100 mg/dl or Impaired Glucose Tolerance (Glucose ≥ 140 mg/dl at 2 hours of 
OGTT) 
b. 
Triglycerides ≥ 150 mg/dl, or on treatment 
c. 
Waist Circumference: Men ≥ 40 in (≥ 102 cm); Women ≥ 35 in (≥ 88 cm) 
d. 
Reduced HDL-C: Men < 40 mg/dl; Women < 50 mg/dl, or on treatment 

19 
 
e. 
Hypertension: ≥130 mmHg systolic, or ≥85 mmHg diastolic, or on treatment 
6. 
HOMA-IR ≥ 2.6.  Our goal is to enroll participants who have pre-existing insulin 
resistance.94 
7. 
hsCRP ≥ 2.0 mg/L. We aim to recruit participants with increased baseline level of 
inflammation. Individuals with hsCRP above 2.0 mg/L have been shown to have an 
increased risk for cardiovascular events.16  
 
VI. 
Exclusion Criteria for subjects with diet-controlled T2DM 
1. 
T2DM that is not well controlled with diet alone: subjects will not be eligible if they take an 
anti-diabetic medication (e.g. metformin, insulin, sulfonylurea, etc.), or have HbA1c > 9%. 
2. 
Presence of a significant active or chronic illness likely to limit life span and/or increase risk 
of intervention, including renal (GFR ≤ 30 ml/min/1.73m2), cardiovascular, hepatic (other 
than obesity-related steatosis), gastrointestinal, immunologic, endocrinologic (e.g. Cushing 
syndrome), pulmonary (other than either asthma not requiring continuous medication or 
sleep apnea-related disorders), or other disorders at the discretion of the investigators. 
3. 
Recent use of colchicine or anorexiant medications in the last 3 months. 
4. 
Known allergy to colchicine. 
5. 
Previous history of agranulocytosis, gout, or significant myositis. 
6. 
Females who are pregnant, planning to become pregnant, currently nursing an infant, or have 
irregular menses, defined as cycles less than 21 days or greater than 45 days.99 
7. 
Individuals who have current substance abuse or a psychiatric disorder or any other 
condition that in the opinion of the investigators would impede competence, compliance, or 
participation in the study. 
8. 
Subjects who regularly use prescription medications unrelated to the complications of 
obesity, especially those known to affect enzymes involved in colchicine metabolism, such 
as CYP3A4 or P-glycoprotein (P-gp) (see Appendix B1).  Oral contraceptive use will be 
permitted, provided the contraceptive has been used for at least two months before starting 
study medication.  The use of over-the-counter and prescription medications will be 
reviewed on a case-by-case basis; depending on the medication, subjects who have 
continued to take prescription medication or have stopped taking an exclusionary medication 
for at least 3 months prior to study entry may be eligible (see Appendix B2). 
9. 
Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight 
change of more than 3% of body weight in the past two months. 

20 
 
10. 
Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the last 
7 days prior to fat biopsy.  
11. 
History of keloid formation. 
12. 
Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes). 
 
 
B. Subject Enrollment 
Subjects will be enrolled only at the National Institutes of Health Hatfield Clinical Research Center. 
Each subject will be registered using the Clinical Research Information System (CRIS), but will be 
considered enrolled only after the participant signs the study consent form.  
 
C. Clinical evaluation and D. Research Testing, and/or Treatment 
Brief summary of study design: Participants who appear qualified when first contacted will 
undergo an initial screening visit, during which a history and physical examination, initial protocol 
review, body composition DEXA, and screening laboratory assessment will take place.  If after the 
screening visit, the subject still appears qualified he/she will return for a baseline visit, during which 
time baseline labs, Hollingshead questionnaire, FSIVGTT, body composition DEXA (if not 
performed during the Screening Visit), CT abdomen, and SAT biopsy will take place. After 
assessments are complete, participants who qualify for and agree to randomization will be 
randomized to either placebo or colchicine and given an eight week supply of medication.  
Randomized subjects will be called at 1 week, 1 month, and 2 months to assess for adverse events 
and adherence.  Randomized subjects will return for a safety interim visit between 4-8 weeks to 
assess for adverse events by physical exam and laboratory studies, where they will receive another 8 
week supply of study drug.  At 3 months, randomized subjects will return for the final visit for 
repeat physical exam, labs, FSIVGTT, body composition DEXA, CT abdomen, SAT biopsy, and 
exit questionnaire.  For randomized subjects who complete the study, they will be given the option 
to return for another “washout” visit three months later, which will involve the same testing as the 
baseline and final visits.  All study visits will take place at the NIH Clinical Center, Bethesda, MD. 
(Please see Appendices A1-A6) 
 
Subject who qualify for the Evaluation-Only arm will only undergo the Screening and 
Baseline visits as described above. 
 
For the Open Label arm, seven subjects with T2DM will be recruited. They will undergo the 
same visits and testing as in the randomization arm, described above.  For the Open Label arm only, 
pills will be dispensed with a “smart bottle cap” which will be able to record the time and date that 

21 
 
the pill bottle was opened. Specifically, we will purchase the MEMSCaps system 
(http://www.mwvaardex.com/index.php/our-adherence-solutions-data-collection/memscap-tm).  
Each bottle supplied by NIH pharmacy will come with a TrackCap®, with the participant’s subject 
ID number assigned to the specific cap.  At the end of the three months on study drug, when the 
subject returns the study drug bottle and cap, we will download the data directly to an SGO lab 
computer.  No PII or medical record number will be associated with the TrackCap, and only the 
investigators involved in the study (e.g. JY, AD, SB) will have access to the data.   
 
i. 
Initial Contact: Telephone Screen 
For participants who respond to advertisements requesting participants (see below), initial 
recruitment and eligibility screening will occur by telephone. Recruitment will occur on a 
continuous basis until study enrollment is complete.  Eligible subjects will then be scheduled for 
an outpatient screening visit.  Eligible subjects will be mailed paperwork in advance, including 
the NIH medical history form, which will then be reviewed in person with the potential subject 
at the outpatient screening visit. 
 
ii. 
Screening Evaluation 
Complete medical history and physical exam will be performed with weight (in kg) 
measured while the subject wears light clothes and height (in cm) measured in triplicate.  
Subjects will rest for 5 minutes in the seated position with the antecubital fossa supported at 
heart level when blood pressure measurements are made. Body mass index (BMI) will be 
calculated before protocol is signed and current medications reviewed, so that individuals taking 
an exclusionary medication, have a disqualifying medical condition, or are weighing more than 
the maximum weight criterion (450 lbs) can be dismissed before the protocol is reviewed.   
Protocol review and signing of consent forms: For those who may be eligible, the relevant 
consent form (either evaluation-only or randomized study protocol) will be discussed and study 
risks and benefits explained. 
EKG and fasting blood for complete blood count (CBC), acute care, mineral, and hepatic 
profiles, PT/PTT, TSH, Free T4, Total T3, IGF-1, and creatine kinase (CK) will be obtained to 
ensure that the subject is in good general health.  Fasting blood samples for hsCRP, insulin, and 
HbA1c will be obtained, and a 2 hour OGTT-GH will be performed to confirm subject eligibility 
for the study.  Additional samples for peripheral blood mononucleated cell (PBMC) analysis and 
future research and genomic DNA isolation will be obtained.   

22 
 
Urine for urinalysis, urine pregnancy test (female participants only), and urine toxicology 
screen will be obtained to ensure that the subject is in good general health and meets protocol 
entry criteria. 
If a potential subject is noted to meet any of the exclusion criteria, that individual will be 
dismissed without further evaluation and referred to their usual physician for any required 
treatment. 
DXA Scan for body composition analysis and fat mass will be obtained.  DXA involves 
passing a collimated X-ray beam of 40 and 70 KeV through the subject and collecting the X-rays 
that pass through the patient with a standard X-ray detector. Because the delivery and attenuation 
of the X-rays are measured separately for the two principal energies, the procedure allows the 
differentiation of substances in their path if the attenuation coefficients of these substances are 
known. Because the delivery and attenuation of the X-rays are measured separately for the two 
principal energies, the procedure allows the differentiation of substances in their path if the 
attenuation coefficients of these substances are known. The manufacturer specifies that the array 
beam delivers an effective dose of 0.00003rem total body irradiation. Participants will undergo 
DXA scans in the Metabolic Clinical Research Unit (MCRU) using the Lunar iDXA (GE 
Healthcare, Madison, WI, USA) machine. The scan takes approximately 20-30 minutes. An 
external standard, simulating bone, fat, and muscle, is scanned once a week to validate 
measurements. This standard allows calculation of total body fat, muscle, and bone mass, and 
percentage of body fat. 
 
iii. 
Baseline evaluation 
An interval history and physical exam will be performed with weight (in kg), height (in cm), 
and blood pressure measured as above. 
Socioeconomic Status will be determined using the Hollingshead Two-Factor Index of 
Social Position.100 
Fasting blood samples of hsCRP, insulin, HbA1c, free fatty acids, lipid panel, uric acid, and 
serum beta HCG (female participants only) will be obtained. Blood samples will also be 
collected for PBMC analysis, as well as to measure inflammatory cytokines, endothelial 
adhesion molecules, lipoproteins (e.g. Lp(a), oxidized HDL, NMR lipoprotein particle analysis), 
leptin, and adiponectin.  Additional research blood samples will be drawn for storage in a -80oC 
freezer for possible future research. 

23 
 
Two stool samples to save for future analyses will be obtained. Subjects will be asked to fill 
out a paper survey including 24 hour food intake that will be reviewed in-person with an NIH 
nutritionist (see Appendix G). 
Urine sample as described previously. In addition to the urine sample collected on 
admission, we will collect a timed eight hour urine sample, which will be saved for future 
analyses.101,102 
FSIVGTT: As described previously,103-105 two angiocatheters will be placed in the 
antecubital veins, one for the administration of glucose and insulin and the other for blood 
sampling from the contralateral arm. A glucose load of 50% dextrose 0.3 g/kg given as a smooth 
bolus over 2 min is given at time 0 and a bolus of 0.05 U/kg insulin is given just before minute 
20. Blood samples are taken at times -15, -10, -5, -1, then at 2, 3, 4, 5, 6, 8, 10, 14, 19, 22, 25, 
30, 40, 50, 70, 100, 140, and 180 minutes after glucose injection for the measurement of plasma 
glucose and serum insulin concentrations.106,107  Insulin administration is essential for individuals 
with severe insulin resistance to achieve a valid test.108-110  Insulin sensitivity and glucose 
effectiveness will be assessed using Bergman’s minimal model,103 which requires the particular 
times of blood collection that have been specified above.  The principal investigator will be 
notified immediately of any glucose measurement less than 40 mg/dL. The principal investigator 
will use his clinical judgment in developing an appropriate, situation-specific management plan. 
 
CT abdomen-adipose:  To quantify the fat content of the liver, subcutaneous, and visceral 
compartments, subjects will undergo a CT of the abdomen, where slices at only three levels will 
be obtained.  CT scanning parameters are 5-10 mm slice thickness, 0.7-0.98 in-plane pixel 
resolution, and 120 kVp. As previously established liver fat can be reliably quantified from one 
slice, typically in the T12-L1 region.111,112 
In accordance with previous studies, subcutaneous and visceral fat compartments can 
reliably be quantified from slices of the L2-3 and L4-5 levels.113  Using automated software 
developed at the NIH, an algorithm consisting of body masking, noise reduction, adipose tissue 
labeling, VAT and SAT separation, and quantification will be used. The body mask is created by 
a region-growing algorithm on the image background that segments the low intensity pixels 
outside the body and in a second pass, removes the CT table. Once the body mask is created, an 
anisotropic diffusion filter reduces noise and voxels between -274 HU and -49 HU. A contour 
around the outside of the body, the “external contour”, is initialized. Active contour models are 
then used to iteratively modify the external contour to find the inner boundary of the SAT; this 
results in a contour along the abdominal wall, specifically the “internal contour”. The adipose 

24 
 
tissue volume inside the internal contour contributes to the VAT and the adipose tissue volume 
between the external contour and internal contour contributes to SAT.   
In collaboration with James Vucich, NIH Medical Physicist, it was determined that the CT 
scan will deliver an effective dose of 0.031 rem total body irradiation per scan.  As randomized 
and open-label participants will undergo three CT scans and three DEXA scans as part of the 
study, the total effective radiation dose for the study is 0.093 rem. Evaluation-only participants 
will receive one CT and one DEXA scan (total effective radition dose = .031 rem. For all three  
protocol arms, this dose is well below the dose guideline of 5 rem per year established by the 
NIH Radiation Safety Committee for adult research subjects. 
Subcutaneous abdominal adipose tissue biopsy: Abdominal subcutaneous adipose tissue 
samples will be obtained using a mini-liposuction technique (see Appendix C for checklist). 
Subjects will be advised not to use non-steroidal anti-inflammatory drugs (NSAID; e.g. aspirin, 
ibuprofen, etc.) for at least one week prior to and following the procedure. The biopsies will be 
performed by an MD or CRNP on the adult outpatient unit. A mixture of 1:1 lactated Ringers or 
normal saline and lidocaine will be used as a local anesthetic until there is an “orange peel” 
appearance to the skin. A sterile approach will be used to minimize risk of infection. Using 10 or 
20 cc syringes with blunt, fenestrated needles, gentle to-and-fro movements will be used to 
obtain the samples. The contents of the syringe will be transferred onto a 250 Nylon mesh (Nitex 
screen cloth, Sefar America, Depew, NY) and rinsed with 5-10ml normal saline to wash away 
the lidocaine and blood clots as much as possible. Fifteen grams of subcutaneous adipose tissue 
samples will be used to assess changes in adipocyte size and the expression of proteins involved 
in inflammation. Subjects will be monitored after the procedure, and post-biopsy care 
instructions will be provided to the subjects. 
 
 
Medication Dispensation and Randomization: Subjects eligible for, and willing to 
undergo randomization will be assigned consecutive study numbers.  Investigators, evaluators, 
and participants will be blinded to assignment to drug or placebo. Assignment of each study 
number to placebo or colchicine will be performed by the NIH pharmaceutical development 
section (before 2017) or the NIH Investigational Drug Management Section, with individuals 
randomized in blocks of four, stratified for sex (male vs. female) and race (black vs. non-black).   
Subjects in the Open Label arm will receive purchased brand-name colchicine (Colcrys).  
  
iv. 
Interim safety evaluations 

25 
 
Randomized and open-label participants will be contacted via telephone to evaluate 
medication tolerability after 1 week and then monthly until they return for their final evaluation. 
If a randomized or open-label participant develops any concerning adverse event, the individual 
will be scheduled for an immediate clinic appointment with a trial investigator for further work 
up, as necessary.  Additionally, randomized and open-label participants will have one interim 
safety visit at the NIH Clinical Center with one of the investigators (e.g. JY, AD, SB) for a 
physical examination and blood work (CBC, HbA1c, acute care profile, hepatic profile, lipid 
panel, insulin, uric acid, hsCRP and CK), between 4-8 weeks.  Samples for PBMC analysis and 
inflammatory cytokines will be collected at this visit.  A urine sample will be obtained as 
described previously. The in-person interim safety visit can take the place of one of the safety 
telephone calls. 
 
v. 
Final evaluation 
For randomized and open-label subjects, an interval history and physical exam will be 
performed with weight (in kg), height (in cm), and blood pressure measured as above.  
An exit questionnaire will be performed (See Appendix D).  
Fasting blood for complete blood count (CBC), acute care, mineral, and hepatic profiles, 
PT/PTT, beta HCG (female participants only), and creatine kinase (CK) will be obtained to 
monitor for adverse events and to ensure that the patient remains in good clinical health. 
Additionally, fasting research blood samples, as described in the baseline evaluation for PBMC 
analysis, inflammatory cytokines, endothelial adhesion molecules, lipoproteins, leptin, 
adiponectin, and research specimens to store for future studies, will be obtained.   
Stool sample for future analyses as described previously. 
Urine sample and eight hour urine collection as described previously.  
FSIVGTT: As described previously. 
DXA Scan as described previously. 
CT Abdomen-for liver and intra-abdominal adipose tissue as discussed previously. 
Subcutaneous abdominal adipose tissue biopsy as described previously. 
 
vi. 
Washout evaluation 
For randomized and open-label subjects, an interval history and physical exam will be 
performed, as above.  
Fasting blood as described in the Final Visit.  
Urine sample and eight hour urine collection as described previously.  

26 
 
Stool sample as described previously. 
FSIVGTT as described previously. 
DXA Scan as described previously. 
CT Abdomen-for liver and intra-abdominal adipose tissue as discussed previously. 
Subcutaneous abdominal adipose tissue biopsy as described previously. 
 
E. Biological Specimens 
i. 
Blood Loss 
a. Screening Visit 
i. Acute care, mineral, hepatic panel, CK,  
8 ml 
TSH, Free T4, T3, insulin, hsCRP, IGF1 
ii. CBC/HbA1c 
3 x2 = 6 ml 
iii. ESR 
1 ml 
iv. PT/PTT 
4.5 ml 
v. OGTT 2-hour glucose, GH 
2 x3 x3= 18 ml 
vi. OGTT hsCRP (at 2hr time point) 
1 mL 
vii. Research (Lavender) 
18 ml 
viii. Research (Green) 
10x4 = 40 ml 
ix. Research (Red) 
20 ml 
x. Research (Red/yellow) 
8 ml 
xi. Research (Gut hormones) 
3 ml 
Total 
127.5 ml 
 
b. Baseline Visit 
i. FSIVGTT 
110 ml 
ii. Glucose (Gray) 
1 ml 
iii. CBC/HbA1c 
3 x2 = 6 ml 
iv. ESR 
1 ml 
v. Acute care, lipid panel, insulin, uric acid, CK 
8 ml 
hsCRP, mineral panel, hepatic panel 
 
vi. Research (Lavender) 
10 ml 
vii. Research (Green) 
10x4= 40ml 
viii. Research (Red) 
10x3= 30 ml 
ix. Research (Red/yellow) 
8 ml 

27 
 
x. Research (for Dr. Sack) 
120 ml (optional) 
xi. Research (Gut hormones) 
3 ml 
Total 
217-337 ml 
 
c. Interim Safety visit (randomized and open-label participants) 
i. CBC/HbA1c 
3 ml 
ii. Acute care, lipid and hepatic panels, CK,  
insulin, uric acid, hsCRP 
8 ml 
iii. Research (Red/yellow) 
8 ml 
iv. Research (Red) 
10 ml 
v. Research (Green) 
10 x3 = 30 ml 
Total 
59 ml 
 
d. Final visit (randomized and open-label participants) 
i. FSIVGTT 
110 ml 
ii. Glucose (GYY) 
1 ml 
iii. CBC/HbA1c 
3 x2 = 6 ml 
iv. ESR 
1 ml 
v. PT/PTT 
4.5 ml 
vi. Acute care, mineral, hepatic, lipid panels, 
8 ml 
insulin, uric acid, CK, hsCRP 
 
vii. Research (Lavender) 
10 ml 
viii. Research (Green) 
10x4= 40ml 
ix. Research (Red) 
10x3 = 30 ml 
x. Research (Red/yellow) 
8 ml 
xi. Research (for Dr. Sack) 
120 ml (optional) 
xii. Research (Gut hormones) 
3 ml 
Total 
221.5-341.5 ml 
 
e. Washout Visit (randomized and open-label participants) 
i. FSIVGTT 
110 ml 
ii. Glucose (GYY) 
1 ml 
iii. CBC/HbA1c 
3 x2 = 6 ml 
iv. ESR 
1 ml 

28 
 
v. PT/PTT 
4.5 ml 
vi. Acute care, mineral, hepatic, and lipid panels, 
8 ml 
insulin, uric acid, CK, hsCRP 
 
vii. Research (Lavender) 
10 ml 
viii. Research (Green) 
10x4=40ml 
ix. Research (Red) 
10x3= 30 ml 
x. Research (Red/yellow) 
8 ml 
xi. Research (for Dr. Sack) 
120 ml (optional) 
xii. Research (Gut hormones) 
3 ml 
Total 
221.5-341.5 ml 
 
Note: Some research blood samples will be sent to our collaborators for analysis of lipid, 
inflammatory, and endothelial parameters.  Other research blood samples will be saved 
for storage in a locked -80oC freezer for possible future research, although there are no 
specific additional studies planned at this time. 
 
ii. Adipose tissue biopsy 
a. Baseline visit 
 
 
 
 
 
 
15 g 
b. Final visit (randomized and open-label participants only)  
15 g 
c. Washout visit (randomized and open-label participants only) 
15 g 
 
 
 
Note: A portion of the adipose samples will be snap frozen and stored in a locked -80oC 
freezer for possible future research, although there are no specific additional studies 
planned at this time. 
 
iii. Urinalysis 
a. Screening visit  
 
 
 
 
10 cc 
b. Baseline visit 
 
 
 
 
 
10 cc + 8hr collection 
c. Interim visit (randomized and open-label participants) 
10 cc 
d. Final visit (randomized and open-label participants) 
10 cc + 8hr collection 
e. Washout visit (randomized and open-label participants) 10 cc + 8hr collection 
 
iv. Stool sample 
a. Baseline visit 
 
 
 
 
 
10 cc 

29 
 
b. Final visit (randomized and open-label participants) 
5 cc 
c. Washout visit (randomized and open-label participants) 5 cc 
 
F. Molecular Genetic Analysis of DNA: Yes.  Samples will be collected and stored for potential whole 
exome or whole genome sequencing for alleles linked to obesity, metabolic syndrome, or response to 
colchicine. 
 
G. Approved Drugs:  
No experimental studies involve approved drugs. Insulin is administered during the FSIVGTT. 
Lidocaine is used during adipose tissue biopsies. 
 
H. Unapproved Drugs: Colchicine is FDA-approved for the treatment of gout and Familial 
Mediterranean Fever. It is unapproved for the amelioration of insulin resistance or metabolic 
syndrome.  In this study we plan to use doses approved and previously proven safe for gout 
prophylaxis – 0.6mg PO BID.58,68  From 2014-2016, the NIH Pharmaceutical Development Section 
used colchicine USP compounding powder to make identical-looking colchicine 0.6 mg and placebo 
capsules. After this date, Pine Pharmaceuticals will follow the same procedures to produce 
colchicine and placebo capsules. For the Open Label arm, participants will receive brand-name 
colchicine (Colcrys).  Participants will be instructed to take one capsule twice daily.  If participants 
develop intractable diarrhea, or other mild side effect that does not warrant medication 
discontinuation and/or study termination, the dose may be reduced to one capsule daily (see Section 
V.A.v. and Appendix F).  If side effects resolve, and the subject remains considerably improved or 
symptom-free for two weeks, the dose may be increased back to twice daily at the investigator’s 
discretion.  All subjects will receive a “Medication Guide” (See Appendix E).  An IND has been 
obtained from the FDA (120722). 
 
I. Sharing of Data with Subjects  
 
Specific results that will be given to participants or their health care providers:  If participants have 
concerning results during any of the visits, this will be communicated both to the patient and their 
designated health care providers. At the conclusion of the entire study, when the randomized 
assignment of subjects to medication is given to the Investigators and the analysis of the results is 
complete, the overall results of the investigation will be given to participants and they will be 
informed of the group to which they were randomized.  For the Open Label arm, once all seven 

30 
 
subjects have completed their participation and analysis of the results of the arm has been completed, 
they will be communicated to the participants of the Open Label arm. 
 
J. Questionnaires/Psychological instruments: The Hollingshead Two Factor Index of Socioeconomic 
Status100 will be administered at the Baseline visit. A non-validated exit questionnaire examining the 
patients’ perceptions of their experience in the protocol will be performed at the final visit. 
 
K. Genetic counseling: n/a 
 
L. Follow-up of participants:  After the baseline visit, randomized and open-label participants will be 
contacted by a study investigator by phone at 1 week, 1 month, and 2 months to assess for any 
adverse outcomes and adherence.  If a participant develops any concerning adverse events, the 
individual will be scheduled for an immediate clinic appointment with a trial investigator for further 
evaluation, as necessary.  Additionally, an interim safety visit at the NIH Clinical Center with one of 
the investigators (e.g. JY, AD, SB) and laboratory examination (e.g. CBC, acute care profile, hepatic 
profile, and CK), will be performed for randomized and open-label subjects between 4-8 weeks. 
 
Participants will be contacted by a study investigator in the event that any abnormal laboratory, 
imaging or EKG findings are found during testing at any of the visits.  At the investigator’s clinical 
discretion in conjunction with the subject’s wishes, these abnormalities may be worked up further at 
the NIH Clinical Center as necessary. 
 
M. Conditions for discharge from the protocol (any of the following):  
a. Completion of the protocol.  Specifically, for evaluation-only participants, the subject 
completes the screening visit and baseline visit.  For randomized and open-label 
participants, the subject also completes at least 90 days of investigational medication 
(colchicine or placebo), and the final visit, as described above. 
b. Development of a significant toxicity, such as myositis, agranulocytosis, acute kidney 
injury (see Section V. A. v.), generally defined as grade II or greater toxicity on the 
CTCAE 4.03 list (see Appendix F and attached file for more information).114 
c. Development of an exclusion criterion, besides T2DM or weight change (see Section IV. 
B.) 
d. Voluntary withdrawal from the trial 
e. Non-compliance with the study procedures at baseline assessment (e.g. unwillingness to 
take investigational medication, undergo FSIVGTT, etc.) 

31 
 
 
V. Analysis of the Study/Oversight 
 
 
A. Clinical Description:   
i. Study Administration and Data Collection, Management, and Storage:   
 
The study will be conducted by the research team as listed above.  Samples and data will 
be coded. Human biospecimens will be collected using procedures appropriate for the 
type of biospecimen being collected and will be handled in accordance with the U.S. 
Occupational Safety and Health Administration’s Bloodborne Pathogens Standard for 
Samples. The principal investigator and the co-investigators will have access to 
identifiable participant records. Data will be coded by subject number, but will be 
readily associated with identifiable participant records. Records containing personal 
identifiers will be maintained consistent with the security measures required by the NIH. 
Data will either be transferred directly from the electronic medical record into the 
NICHD Clinical Trials Data Base (CTDB), or for data where direct transfer is 
unavailable, will be double-entered by hand into CTDB.  All data will be kept on secure 
computers in the SGO office, whose doors will be locked when staff is not present.  All 
paper data will also be kept in locked areas in the SGO office. 
Serum and plasma samples not analyzed immediately (as described above) will be stored 
in locked freezers by the NICHD sample management team supervised by the NICHD 
Clinical Director and tracked/managed through CTDB. Human biospecimens in storage 
will have a unique identifier, will be labeled with a printed label and will contain a 
barcode. Samples will be tracked using a computer‐based inventory system that records 
the location and detailed information of every specimen in the repository. 
Sample freezers will be operated using facility environments that include ambient 
temperature controls, good air circulation, lighting, and security. Systems are in place to 
allow for local and remote temperature monitoring of freezers. The NIH has an 
emergency preparedness plan that covers equipment failures and power interruption, 
including back‐up storage capacity and back‐up power generators.  
 
Participants give explicit permission regarding sample use in the consent form of the 
study, indicating if samples may be used for future experimental testing related to 
colchicine response, body weight and metabolism and specify if additional consent is 

32 
 
required.  At the conclusion of the study, samples will be retained for future 
experimental testing related to colchicine response, body weight and metabolism. 
 
The PI will inform the IRB if obtained samples are lost or destroyed before they are 
assayed for primary or secondary outcomes. Loss or destruction of samples saved only 
for tertiary outcomes or future unspecified research will not be routinely reported, unless 
such loss also entails a breach of the Privacy Act. 
 
Data safety monitoring board (DSMB): A data safety and monitoring board will be 
assembled as required by NICHD for pharmaceutical studies. 
 
ii. Publication of Research Findings:   
 
On conclusion of the randomized trial, it is hoped that the results of this pilot 
investigation will be publishable. If so, findings will be submitted to a peer-reviewed 
journal. Regardless of publishability, results will be deposited on www.clinicaltrials.gov 
as required by governmental rules. 
 
iii. Statistical Analysis of the Data 
 
Data analyses will be conducted by the investigators. The primary outcome of the 
randomized study will be change in insulin sensitivity as calculated by the minimal model 
from data obtained through the FSIVGTT in subjects who are randomized to colchicine 
versus those given placebo. Secondary outcomes will be changes in metabolic parameters 
(e.g. AIRg, disposition index, glucose effectiveness, HOMA-IR, HbA1c, lipid panel), and 
the inflammatory marker hsCRP.  Additionally, changes in other inflammatory markers, 
endothelial cell adhesion markers, PBMC inflammatory parameters, SAT adipocyte size, 
SAT inflammatory parameters, and adipose tissue compartments will be evaluated as tertiary 
outcomes.   
 
All analyses will be planned to be performed as intention-to-treat for all randomized 
subjects.  For the primary outcome and the secondary outcomes, we will use an analysis of 
covariance (ANCOVA) to examine the differences between treatment arms, taking into 
account age, baseline body fat percentage, change in body fat percentage, and sex. Two-
sided significance tests will be performed for these analyses, so that both positive and 
negative treatment effects may be detected.  Spearman rank will be used for correlations. 

33 
 
 
For the open-label trial of seven patients with T2DM, the mean change (and the 
standard deviation) for the same variables pre-post treatment will be evaluated to help 
calculate the power of a potential future RCT. 
 
iv. Monitoring Subjects and Criteria for Withdrawal 
 
Screening evaluations may lead to withdrawal of subjects who are not eligible according to 
inclusion/exclusion criteria. Noncompliance with study procedures before subjects are started 
on drug may lead to withdrawal of study participants. Subjects will be followed using an 
“intention to treat” model once randomized.  Phone contacts will be made to review 
medication tolerance and assess for development of serious adverse events after 1 week, 1 
month, and 2 months of treatment. Subjects will be seen if serious adverse events are 
suspected. If subjects are seen by their own physicians for illnesses, we will request the records 
for those visits to determine if they are possibly related to the research. 
 
v. Stopping Rules: 
 
Events that will result in discontinuation of treatment include: pregnancy or serious 
unanticipated complications, including development of any medical problem listed in the 
exclusion criteria after starting on drug. Subjects will be excluded if they have any verified 
abnormality in laboratory values or physical status that cannot be explained by a mild 
intercurrent illness (or by obesity and insulin resistance itself) that in general are considered  
CTCAE grade 2 (See Appendix F) or greater for gastrointestinal, hematologic, clinical 
hemorrhage, kidney/bladder, alopecia, pulmonary, cardiac, neurocerebellar, allergy, flu-like 
symptoms, metabolic, or eye toxicities; and grade 3 or greater for infection, anorexia, 
circulatory, neurologic, dermatologic, coagulation, endocrine, or performance status 
toxicities.115  Problem-specific stopping rules have been defined in the CTCAE v4.03 
submitted to the IRB in conjunction with this protocol.  Investigator error leading to initiation 
of treatment in an ineligible subject will lead to treatment discontinuation as soon as the error 
is uncovered. Should discontinuation be required for any reason, no tapering of colchicine is 
necessary; subjects will be requested to return all unused medication.  Because the randomized 
study is a pilot intended to identify the sample size needed for a fully-powered trial, there are 
no formal stopping rules that would halt the study before all participants have been 
randomized. 

34 
 
 
B. Molecular Studies 
 
Samples will be stored for future genetic studies of variants associated with body weight and 
inflammation. Examples of genes that may be examined include obesity-associated genes such 
as leptin, leptin receptor, melanocortin 4 receptor, melanocortin 3 receptor, brain-derived 
neurotrophic factor, the beta 3 adrenergic receptor, and SIM1, as well as inflammatory genes 
such as NLRP3, ASC, procaspase-1, IL-1, and IL-6. The consent form has been prepared to 
allow participants agree to whole-exome/genome sequencing for identification of genetic 
changes linked to inflammation, obesity or obesity-linked dysregulation of metabolism. 
 
In some instances, investigators will share coded specimens from this protocol with 
collaborators who will not be provided with a key to the code.  Subcutaneous adipose biopsy 
specimens will be sent to Dr. Vincent Manganiello’s lab in a de-identified manner, where they 
will be examined for local inflammation.  Dr. Joshua Zimmerberg’s lab will receive coded 
samples for analysis of inflammation and metabolic function, such as adipose GLUT4 mobility.   
 
In collaboration with the Center for Human Immunology, Autoimmunity, and 
Inflammation (CHI) at the NIH, coded samples will be sent for PBMC analysis, including 
mRNA expression array analysis, flow cytometry and cryopreservation.  Samples of PBMC for 
protein lysates will be analyzed by Dr. Vincent Manganiello’s lab via Western Blot analysis to 
examine colchicine’s effects on NLPR3 inflammasome assembly, as well as for its effects on 
inflammasome-mediated IL-1B production.   Dr. Michael Sack, NHLBI, will also receive coded 
serum samples for analysis of PBMC inflammation. 
 
Dr. James Goedert (NCI) will receive coded samples of blood for analysis of estrogen 
and androgen markers, as they may change with changes in insulin sensitivity. For collaborators 
outside the NIH, The PI will establish these collaborations through an OHSRP request for 
determination and provide a list of all collaborations established in this manner to the IRB at the 
time of CR. 
 
Stool samples will be obtained for future analyses to be performed by CHI. 
 
Dr. Perry Blackshear, NIEHS, will received coded samples of blood and adipose tissue 
for analysis. 
 
VI. Determination of Sample Size:  
 

35 
 
 
The randomized trial is a pilot study, intended to determine the treatment effect size and standard 
deviation for colchicine’s effects on change in insulin sensitivity, to allow for adequate powering of a larger 
future study.  Nonetheless, we will power it so it might be able to detect a moderate-to-large difference 
between treatment groups.  Based on the data of prior studies,116,117 the standard deviation (SD) for change in 
insulin sensitivity as measured by FSIVGTT is about 2 mIU/L/min.  If a similar SD is found in the present 
study, we estimate that a total sample size of 40 subjects (20 colchicine and 20 placebo) is necessary to have 
80% power (β) to detect a 60% difference in insulin sensitivity between colchicine and placebo groups, at 
level of significance <0.05 (2-sided). 
  
VII. Human Subjects Protection: 
A. Rationale for Subject Selection:  This study seeks to understand how colchicine affects insulin 
sensitivity, pancreatic beta-cell function, and low-grade inflammation in humans.  Because obese 
participants have a greater level of insulin resistance than non-obese individuals, and participants 
with MetS have a greater level of inflammation than those without, we selected to test obese 
individuals with metabolic syndrome who demonstrate both high insulin resistance and evidence 
for inflammation.  Participants with type 2 diabetes mellitus frequently are taking medications 
that may confound the results of the tests we plan to study.  Additionally, diabetics are presumed 
to have beta-cell destruction which may be too far advanced to demonstrate benefit from an 
intervention, whereas a disease process in an earlier stage (e.g. insulin resistance and/or 
prediabetes) may demonstrate benefit. On the other hand, diabetics also have the greatest level of 
obesity-associated inflammation, so they may demonstrate benefit from colchicine.  To 
investigate this further, we will recruit seven subjects with diet-controlled DM to an Open Label 
arm.    
In order to expand our knowledge of local insulin action at the level of the adipocyte, we 
also plan to enroll subjects across a larger spectrum of insulin sensitivity/resistance for testing 
only at the Screen and Baseline Visits.  Subjects who are not eligible to be randomized to study 
drug but who will be enrolled to the Evaluation Only arm will include those who are not obese, 
obese but without significant insulin-resistance or evidence of inflammation, and those with diet-
controlled type 2 diabetes. Such subjects are excluded, as listed above, only if they are pregnant 
or nursing, use medications or substances that affect the validity of measures, have a significant 
medical/psychiatric condition, or weigh too much for required procedures. 
Children are not studied because the nature of the experiment involves a minor increment 
above minimal risk (adipose tissue biopsies, potentially toxic medication), that can be answered 
in adult subjects.   

36 
 
Adults of all races and ethnicities will be included. Since metabolic syndrome is identified in 
large percentages among minority groups including Asian Americans, African American and 
Hispanic Americans, we anticipate good representation of all major races/ethnicities. It is 
expected, given our recruitment procedures targeting individuals living in the greater 
metropolitan Washington D.C. area, that we will recruit rates of minority participation in excess 
of their representation in the Washington, D.C. metropolitan area: 64.2% Caucasian, 27.2% 
African-American and 8.6% Hispanic. 
B. Participation of Children and Other Vulnerable Populations: n/a 
C. Screening Methodology 
Subjects who appear eligible when assessed during a phone screening intended to assess general 
health and body weight/height (to determine BMI) will attend a screening visit at the NIH 
Clinical Center, during which time eligibility will be determined. History and physical 
examination will be conducted by a licensed practitioner. Laboratory data from blood draw and 
pregnancy testing will be reviewed before subjects are scheduled for the Baseline visit and 
randomization to drug/placebo (or initiation of colchicine in the open-label arm).  
 
Information regarding the key assays used for eligibility criteria (all measured by the NIH 
Clinical Center Laboratory Medicine Department): 
 
Glucose 
 @75mg/dL    
: intraassay CV 2%             interassay CV 3% 
 @ 379 mg/dL  
: intraassay CV 1%             interassay CV 2% 
Sensitivity: 1 mg/dL 
 
  
Insulin 
@ 24.2 µU/mL    
: intraassay CV 1.2% 
 
interassay CV 6.0% 
@ 82 µU/mL  
: intraassay CV 1.0% 
 
interassay CV 2.5% 
Sensitivity: 0.2uU/mL 
  
hsCRP 
@ 0.239 mg/dL     : intraassay CV 5.2%              
interassay   5.2% 
@ 0.650 mg/dL    
: intraassay CV 5.0%             
interassay   5.7% 
Sensitivity: 0.016 mg/dL 

37 
 
  
Triglycerides 
@ 68 mg/dL     
: intraassay CV 3%            
interassay CV 4% 
@384 mg/dL    
: intraassay CV 2%            
interassay CV 2% 
Sensitivity: 2 mg/dL 
  
HDL cholesterol 
@ 26 mg/dL    
: intraassay CV 2.3%            
interassay CV 2.7%   
@ 47 mg/dL    
: intraassay CV 1.6%            
interassay CV 2.3%   
@ 67 mg /dL   
: intraassay CV 1.9%           
interassay CV 2.1% 
Sensitivity: 3 mg/dL 
 
Hemoglobin A1c 
@ 5.1% 
 
: intraassay CV 0.4% 
 
interassay 1.11% 
@9.9% 
 
: intraassay CV 0.5% 
 
interassay 0.73% 
Sensitivity: 4.2% 
 
D. Risks/Benefits Analysis (including considerations of alternatives to participation) 
1. Potential Benefits, in addition to have the results of all screening testing, (as outlined above) 
from randomization include the following 
a. Improving insulin sensitivity 
b. Improving pancreatic beta-cell reserve 
c. Decreasing inflammation 
d. Decreasing cardiovascular risk 
e. Benefit to medicine and society 
2. Risks/Discomforts 
a.    Colchicine (randomized and open-label participants only) 
i. Adverse Reactions 
Among post-marketing experience in the use of Colcrys (the trade name for 
colchicine that is available in the US), adverse reactions have been generally 
reversible upon temporarily interrupting treatment or lowering the dose of 
colchicine. The following adverse reactions have been reported with colchicine: 
- 
Neurological: sensory motor neuropathy 
- 
Dermatological: alopecia, maculopapular rash, purpura, rash 

38 
 
- 
Digestive: abdominal cramping, abdominal pain, diarrhea, lactose intolerance, 
nausea, vomiting 
- 
Hematological: leukopenia, granulocytopenia, thrombocytopenia, 
pancytopenia, aplastic anemia 
- 
Hepatobiliary: elevated AST, elevated ALT 
- 
Musculoskeletal: myopathy, elevated CPK, myotonia, muscle weakness, 
muscle pain, rhabdomyolysis 
- 
Reproductive: azoospermia, oligospermia  
Tolerance, abuse, or dependence with colchicine has not been reported.68 
 
In recent clinical trials, the most commonly reported adverse reactions were 
abdominal symptoms such as diarrhea, cramping, nausea, diarrhea, abdominal pain, 
and vomiting. For example, in a randomized, placebo-controlled trial investigating 
colchicine 0.6mg PO bid for 6 months in participants after coronary angioplasty, of 
the 130 subjects in the intervention group 28% developed diarrhea, whereas other 
side effects, including nausea or rash, were not significantly different from 
placebo.118  Another randomized placebo-controlled study investigating colchicine 
0.5mg PO bid for 6 months in DM participants undergoing  percutaneous coronary 
intervention (PCI) with a bare metal stent (BMS), of the 196 subjects 16% 
developed diarrhea or nausea in the colchicine group vs. 7% in the placebo group.81 
 
Subjects who may develop renal or hepatic impairment due to causes independent of 
participating in the protocol are at increased risk for developing adverse reactions 
and toxicities as colchicine is metabolized in the liver, and excreted in significant 
amounts in the bile and urine. (See Section VII. H.) 
 
ii. Fatal Overdose 
Fatal overdoses, both accidental and intentional, have been reported in adults and 
children who have ingested colchicine. The exact dose of colchicine that produces 
significant toxicity is unknown. Fatalities have occurred after ingestion of a dose as 
low as 7 mg over a 4-day period, while other participants have survived after 
ingesting more than 60 mg. A review of 150 participants who overdosed on 
colchicine found that those who ingested less than 0.5 mg/kg survived and tended to 
have milder toxicities, such as gastrointestinal symptoms, whereas those who took 

39 
 
0.5 to 0.8 mg/kg had more severe reactions, such as myelosuppression. There was 
100% mortality in those who ingested more than 0.8 mg/kg.  
 
The first stage of acute colchicine toxicity typically begins within 24 hours of 
ingestion and includes gastrointestinal symptoms, such as abdominal pain, nausea, 
vomiting, diarrhea, and significant fluid loss, leading to volume depletion. 
Peripheral leukocytosis may also be seen. Life-threatening complications typically 
occur 24 to 72 hours after drug administration, and death may result from respiratory 
depression and cardiovascular collapse. Individuals who survive a severe overdose 
may develop rebound leukocytosis and alopecia one week after the initial ingestion. 
Treatment of colchicine poisoning should begin with gastric lavage and supportive 
measures. No specific antidote is known, and colchicine is not effectively removed 
by dialysis. 
 
 
 
 
iii. Drug Interactions 
Colchicine is metabolized by P-gp and CYP3A4. Life-threatening and fatal drug 
interactions have been reported in individuals taking colchicine concurrently with P-
gp and/or strong CYP3A4 inhibitors (see Appendix B1). If treatment with a P-gp or 
strong CYP3A4 inhibitor is required in a subject, the investigational medication will 
be discontinued and the patient will be withdrawn from the protocol.  
 
iv. Pregnancy 
Adequate well-controlled studies on the use of colchicine in pregnancy do not exist.  
Among published reports of pregnant women using colchicine to control their 
Familial Mediterranean Fever, no increased risk of miscarriage, stillbirth, or 
teratogenic effects was seen.  However, colchicine did demonstrate fetal toxicity and 
teratogenicity in the offspring of pregnant rats.  Subjects currently pregnant, 
breastfeeding, or planning to become pregnant will be excluded from the study.  
Enrolled female subjects will be strongly advised to use contraception and condoms 
if sexually active. As noted above, women who become pregnant while taking study 
medication will be immediately told to stop medication and will be withdrawn from 
all study procedures. The study team will follow up with the participant regarding 
the outcome of the pregnancy.  
 

40 
 
b. Physical Examination by a health care provider involves no known risk, but does take 
time. A limited physical examination will be performed in a concise but thorough 
manner with appropriate measures taken to provide the privacy of the subject. 
c. Questionnaires are also without significant risk, but are inconvenient because of the 
time required to complete. 
d. DXA scanning is not painful, but may be inconvenient because of the time needed to 
complete the study. DXA scanning involves radiation exposure for research purposes 
only.  According to the manufacturer’s specifications, DXA scanning for body 
composition using the array beam delivers an effective dose of 0.00003rem total body 
radiation. This calculated value is known as the “effective dose” and is used to relate the 
dose received by each organ to a single value. This amount of radiation from the three 
DXA scans to be performed in this study is 0.00009 rem. For comparison, the average 
person in the U.S. receives a radiation exposure of 0.3 rem/yr from natural background 
sources, such as from the sun, outer space, and from radioactive materials that are found 
naturally in the earth’s air and soil.  
e. Blood withdrawals will remain well within the NIH guideline for adults of 7 ml/kg 
every 6 weeks. Blood collection with venipuncture is associated with mild discomfort 
and the possibility of localized bruising or extravasations. The risk of infection, 
phlebitis, or fainting is extremely small. 
f. Stool samples are also without significant risk, but some subjects may find them 
uncomfortable to collect.  The surveys and food records are without significant risk, but 
may be inconvenient because of the time required to complete. 
g. Oral glucose tolerance test is also is without significant risk, but is inconvenient 
because of the time required to complete the study. The only risks from this kind of 
study are those described for any blood draw, as well as developing nausea from the oral 
glucose.  As we are screening out patients with diabetes mellitus, the risk of developing 
hyperglycemia exists, but in most cases should be transient and not clinically significant. 
h. Subcutaneous abdominal adipose tissue biopsies. The risks associated with biopsies 
include pain, bleeding, bruising, infection, numbness related to nerve injury, and 
scarring or keloid formation. Lidocaine will be used to diminish pain. A sterile approach 
will be incorporated to avoid risk of infection, and to our knowledge, there has not yet 
been a report of infection. After the procedure, sterile dressing will be applied, in 
addition to an ice pack; the latter will be used to decrease bleeding/hematoma formation. 
In the immediate post-procedure period (while subjects are observed), the site will be 

41 
 
monitored for immediate complications, and addressed appropriately. Biopsies from 
abdominal subcutaneous fat will be taken. Subjects will also be instructed on signs of 
these risks and to observe the site after discharge, and to immediately report any 
concerns to the clinical team. The physician/nurse practitioner who is in charge of 
performing the biopsy will fill out a "Surgical/Invasive Procedure Verification 
Checklist” during the procedure to make sure every prerequisite to ensure safety of the 
subject is met. If increased occurrences of any of these risks are noted, technique will be 
adjusted in attempt to reduce the risk. 
i. 
Frequently Sampled Intravenous Glucose Tolerance Test (FSIVGTT) is 
inconvenient because of the time it takes to complete, the need for two intravenous lines, 
and the need to be confined to bed.  The only risks from this kind of study are those 
described for any blood draw (see #5), mild arm tenderness while the dextrose is 
infused, and the possibility of a decreased blood glucose value from the administration 
of insulin. The standard dose of insulin administered for evaluation of growth hormone 
secretory capacity, 0.05 U/Kg, will be administered.  With mildly low blood sugars (< 
50 mg/dL), subjects get hungry and may become drowsy or sweaty.  With blood sugars 
under 30 mg/dL, subjects may be difficult to arouse or have seizures.  However, because 
the obese adults to be studied on this protocol will all have some degree of insulin 
resistance, significant hypoglycemia is not anticipated.  Nevertheless, a physician or 
nurse practitioner will be present during this test for the first hour of the test (including 
the first thirty minutes following insulin administration) to monitor for signs of 
significant hypoglycemia. The principal investigator will be notified immediately of any 
glucose measurement less than 40 mg/dL. The principal investigator will use his clinical 
judgment in developing an appropriate, situation-specific management plan.  Treatment 
will take place as needed with dextrose or glucagon according to clinical evaluation. 
j. 
CT Abdomen for hepatic and intra-abdominal adipose tissue is not painful, but may 
be inconvenient because of the time needed to complete the study.  Some people feel 
anxious while inside the CT scanner. 
 
Computed tomography involves radiation exposure for research purposes only.  
Although each organ will receive a different dose, the amount of radiation exposure 
subjects will receive per scan is equal to a uniform whole-body exposure of 0.031 rem. 
This calculated value is known as the “effective dose” and is used to relate the dose 
received by each organ to a single value. The total effective radiation dose that patients 
will be exposed to in the study is 0.093 rem, well within the dose guideline established 

42 
 
by the NIH Radiation Safety Committee for adult research subjects. The guideline is an 
effective dose of less than 5 rem/yr.  No contrast dye will be administered as part of this 
protocol. 
 
One possible effect that could occur at these doses is a slight increase in the risk of 
cancer. The natural chance of a person getting a fatal cancer during his/her lifetime is 
about 25 %. The increase in the chance of getting a fatal cancer as a result of the 
radiation exposure received from this research study is 0.1%. Therefore, the total risk of 
fatal cancer may be estimated to increase from 25% to 25.1%. This change in risk is 
small and cannot be measured directly. Compared with other everyday risks, such as 
flying in an airplane or driving a car, this increase is considered slight. 
 
Subjects will be advised to notify the research staff if they have participated in 
research studies at the NIH or other institutions that involved the use of radiation so as to 
make sure that the total radiation dose from all studies is not excessive. Women with 
childbearing potential must have a negative pregnancy test before undergoing the CT 
scan. 
k. “Smart” Pill Bottle Cap.   We will use  a “MEMS cap” (MWV Switzerland, Ltd) on 
the top of the medicine bottles to record patient adherence to the study drug. As 
described above, the cap records the date and time that the bottle is opened, and the data 
will be downloaded by one of the protocol investigators onto a laboratory computer.  
This will pose no additional risk to the subjects.    
 
l. 
Testing for DNA including exome/whole genome sequencing. The risks from DNA 
tests are primarily the risk of discovery of incidental medical information, risks of 
genetic discrimination and risks related to confidentiality and data availability.  
a. 
Incidental medical information.  Genetic risk factors may be found for 
diseases unrelated to the condition for which the patient is being studied. This risk is 
inherent in any genetic analysis; however, the risk increases with increased 
magnitude of data and degree of analysis.  This study will uncover incidental 
information. In some cases this information will provide additional details about 
diseases already known to be present in a family. For example, identification of a 
BRCA1 mutation may be new information for a family, but they may already know 
that breast cancer occurs in their family. Similarly, identification of a Huntington 
disease mutation is not likely to occur in a family without any prior family history. 
Clinically significant information could be identified for diseases that were not 

43 
 
previously recognized to be a concern in a given family.  This could occur with 
dominant disorders that show decreased penetrance or with recessive genetic 
disorders. Exome studies can detect carrier status for recessive disorders such as 
cystic fibrosis or sickle cell anemia.  Determination of carrier status for recessive 
disorders will not impact the subject’s health directly, but this information does have 
potential reproductive implications for the subject and other members of the family. 
This information could cause anxiety in some individuals; however, being aware of 
these conditions provides an opportunity for genetic counseling. 
As genetic understanding of genetic risk factors for multifactorial disorders 
increases, we may detect vast numbers of polymorphisms that slightly increase the risk 
of a specific disorder such as diabetes or heart disease.  The knowledge of these risk 
factors could provide an opportunity to alter lifestyle factors that contribute to the risk, 
but at the same time could produce anxiety.  Unless felt to be of urgent medical 
importance, we will not report these results to patients or families.  General principles 
guiding a decision to inform guardians about genetic information obtained from this 
study will be: 
i. The genetic change must be known or predicted to be of urgent clinical 
significance to either a proband or a first degree relative. 
ii. Knowledge of the finding must have a clear direct benefit that would be lost if 
diagnosis was made at a later time point.  Specifically, knowledge of this risk 
factor will substantially alter the patient’s medical care.   
iii. The potential benefit of knowing a genetic disorder exists clearly outweighs the 
potential risks of anxiety and subsequent medical testing that could result from 
this knowledge. 
iv. Autosomal recessive mutations will only be reported if a) the carrier frequency 
for mutations in that specific gene is greater than 1% (This corresponds to 
disorders with a disease incidence of more than 1/40,000), b) the syndrome results 
in significant morbidity, c) early diagnosis and intervention would have 
significant benefit for an affected individual, and d) early diagnosis is not likely 
to be made by other means.   
 
The decision to notify or not notify will be made by the study investigators based on the 
state of genetic knowledge at the time of data analysis.  Although not precluded, the 
database will not be actively screened for changes that in the future are found to be 

44 
 
associated with a specific disease.  If a genetic change of clinical significance is 
detected, subjects will be given the option to have confirmatory testing performed in a 
CLIA approved laboratory.  Confirmatory testing will be performed on new DNA 
samples obtained from the probands.  Identification of genetic disease in a proband 
could produce anxiety for the parent, siblings or other relatives.  Relatives of the proband 
could learn that they are at risk for a genetic disorder. 
 
b. Genetic Discrimination:  If an individual elects to have a genetic finding 
confirmed, that information will then become part of the medical record at the NIH.  
This information will be released to third parties if the individual sign a release of 
information form.  An insurance company or an employer could interpret this to be a 
pre-existing condition to deny benefits or coverage.   This however, is largely a 
theoretical concern.  No cases of discrimination are known to have resulted due to 
participation in a research study. The risk of genetic discrimination in the context of 
health insurance and employment should also be decreased by Genetic Information 
Nondiscrimination Act of 2008. 
c. Confidentiality and Data availability:  Samples and data will be coded.  Only 
individuals involved in consenting and patient care will have access to identifiers.  
Records containing personal identifiers will be maintained consistent with the security 
measures required by the NIH. We will follow guidelines developed by the NIH for 
projects funded by the Federal Government.  In lieu of specific guidelines, subjects 
may opt out of having sequence data deposited in public databases.  
 
Results of genetic tests performed for research will not be placed in the medical record.  
It is possible that we will send subject’s DNA to other scientists working with us on 
obesity gene studies, but in this case personal identifiers will be removed. If a subject 
chooses to withdraw from the study, we will keep the DNA for analysis, unless the 
subject requests that the sample be destroyed. 
 
3. Assessment of Risk/Benefit Ratio:  Although it is impossible to quantify the exact risk to 
benefit ratio, we feel that the risks as mentioned above are a minor increment above minimal 
risk and mitigated whenever possible.  The potential benefit to the randomized and open-
label subjects can be large with a decrease in cardiovascular risk factors, improvement in 
metabolic profile, and decrease in chronic inflammation.  Additionally, we feel that this 

45 
 
study will contribute significant knowledge to the field of medicine in examining a novel 
therapeutic approach in humans to improve insulin resistance and metabolic syndrome.  To 
date the effects of disrupting the NLRP3 inflammasome on metabolic parameters and 
glucose homeostasis have not been studied in humans with metabolic syndrome. 
 
4. Alternatives to participation:  The major alternative is not to participate in this experiment, 
as there are no inflammasome-reducing medications approved for patients with metabolic 
syndrome. Participants could also consider starting a formal lifestyle modification program, 
including improved diet and exercise.  Established medical options for obesity and metabolic 
syndrome include undergoing bariatric surgery or starting weight-loss medications (e.g. 
lorcaserin or phentermine/topiramate).  Participants can look up research alternatives for 
interventions for obesity or metabolic syndrome on www.clinicaltrials.gov.     
 
E. Privacy and Confidentiality Provisions 
We will follow all relevant NIH policies and HRPP SOPs. Samples and data will be coded.  
Only individuals involved in consenting and patient care will have access to identifiers.  Records 
containing personal identifiers will be maintained consistent with the security measures required 
by the NIH. Access to the code will be limited to the study investigators (e.g. JY, AD, TC, and 
SB).  Electronic data will be password protected and accessible only by authorized study 
personnel. Electronic data will be backed up daily and stored at a secure server. All hardcopy 
data will be locked in Dr. Yanovski’s storage cabinets in research offices at the NIH Clinical 
Research Center and will be accessible only by authorized study personnel. All data will be 
collected specifically for the purpose of the current proposed research project. Any email 
communications that involve PII will be sent using encryption or the secure email system of the 
NIH. Since even when precautions are taken to ensure participant confidentiality, data collection 
in any study is accompanied by threat to privacy and confidentiality, in the unlikely event of a 
potential breach of confidentiality, the PI will immediately inform the IRB. The hsCRP and other 
clinical chemistry results are done by NIH Clinical Pathology and will be in the medical record. 
Results of the serum cytokine profile, endothelial adhesion profile, lipoprotein NMR, DXA, 
DNA, and adipose tissue studies will not be placed in the medical record.   
 
In the Clinical Trials Database, clinical and demographic information will include age; sex; 
race/ethnicity; family history of diabetes, hypertension, or hyperlipidemia and results entered 
from each case report form for visits and phone conversations. 

46 
 
 
F. Compensation 
We will use NIH guidelines for payment of research volunteer. The remuneration is based on 
time and inconvenience during the stay. Payment will be as follows: 
 
[For randomized or open-label subjects] 
1. Outpatient screening visit: 
$100 
2. Baseline evaluation visit: 
$300 
3. Interim Safety Visit: 
$50 
4. Final Visit: 
$300 
5. Washout Visit: 
$300 
       Total= 
$1050 
 
[For evaluation-only subjects] 
1. Outpatient screening visit: 
$100 
2. Baseline evaluation visit: 
$300 
 
Total= 
$400 
 
 
G. Consent Procedures 
Each subject will receive a written explanation of the purposes, procedures, and potential 
hazards of the study (See Consent Form). The purpose of the project, all testing procedures, and 
study components will be described in detail. Potential participants will also be informed of the 
possible risks and inconveniences of the study. Consent will be obtained only by the principal 
and associate investigators and Key Research Personnel designated on the first page of the 
protocol (e.g. JY, AD, SB), in person at the beginning of the first Screening Visit.  
Communication of this information and of the subject’s consent will be documented in the 
medical record. All subjects will be informed of their right to withdraw from the study at any 
point of time during the study. 
 
Non-English–Speaking Participants 
If a non-English speaking participant is unexpectedly eligible for enrollment, the participant 
will be provided with the CC Short Written Consent Form for Non-English Speaking Research 
Participants in the participant’s native language and a verbal explanation of the purpose, 

47 
 
procedures and risks of the study as described in MAS Policy M77-2, NIH HRPP SOP 12 and 
45 CFR 46.117(b)(2). The IRB-approved English consent form will serve as basis for the verbal 
explanation of the study. The investigator will obtain an interpreter unless the investigator is 
fluent in the prospective participant’s language. Preferably, the interpreter will be someone who 
is independent of the participant (i.e., not a family member). Interpreters provided by the CC 
will be used whenever possible. The interpreters will interpret all oral communications (English 
to target language and conversely) about the IRB-approved English consent form, facilitate 
discussion between the participant and investigator, and ensure understanding. 
The IRB-approved English consent form will be signed by the investigator obtaining consent 
and a witness to the oral presentation. The CC Short Written Consent Form will be signed by 
the participant and a witness who observed the presentation of information. The interpreter may 
sign the consent document as the witness and, in this case, will note “Interpreter” under the 
signature line. A copy of both signed forms will be provided to the participant to take home. 
The investigator obtaining consent will document the consent process in the participant’s 
medical record, including the name of the interpreter.  
 
We request prospective IRB approval of the use of the short form for up to a maximum of 5 
separate encounters in a given language and will notify the IRB at the time of continuing review 
of the frequency of the use of the Short Form. Should we reach the threshold of 5, we will 
notify the IRB of the need for an additional use of the Short Form and that we will have that 
consent document translated into the given inherent language. 
 
H. Pharmaceutical Information 
Colchicine is an alkaloid chemically described as (S)N- (5,6,7,9-tetrahydro- 1,2,3,10-
tetramethoxy-9-oxobenzo [alpha] heptalen-7-yl) acetamide with a molecular formula of 
C22H25NO6 and a molecular weight of 399.4. 
 
Pharmacokinetics 
Absorption: “In healthy adults, colchicine is absorbed when given orally, reaching a mean Cmax 
of 2.5 ng/mL (range 1.1 to 4.4 ng/mL) in 1 to 2 hours (range 0.5 to 3 hours) after a single dose of 
0.6 mg administered under fasting conditions.  After 10 days on a regimen of 0.6 mg twice daily, 
peak concentrations are 3.1 to 3.6 ng/mL (range 1.6 to 6.0 ng/mL), occurring 1.3 to 1.4 hours 
post-dose (range 0.5 to 3.0 hours). In some subjects, secondary colchicine peaks are seen, 
occurring between 3 and 36 hours post-dose and ranging from 39% to 155% of the height of the 

48 
 
initial peak. These observations are attributed to intestinal secretion and reabsorption and/or 
biliary recirculation.  Absolute bioavailability is reported to be approximately 45%. 
Administration with food has no effect on the rate of absorption, but does decrease the extent of 
colchicine by approximately 15%, which is likely clinically insignificant.”68  
 
Distribution 
“The mean apparent volume of distribution in healthy young volunteers was approximately 5 to 
8 L/kg. Colchicine binding to serum protein is low, 39 ± 5%, primarily to albumin regardless of 
concentration”68.  Accumulation into neutrophils and macrophages is slow, only reaching peak 
concentration at 48 hours.119  
 
Colchicine crosses the placenta (plasma levels in the fetus are reported to be approximately 15% 
of the maternal concentration). Colchicine also distributes into breast milk at concentrations 
similar to those found in the maternal serum.68 
 
Metabolism 
Colchicine is demethylated to two primary metabolites, 2-O-demethylcolchicine and 3-O-
demethylcolchicine (2- and 3-DMC, respectively), and one minor metabolite, 10-O-
demethylcolchicine (also known as colchiceine), primarily through CYP3A4. Plasma levels of 
these metabolites are minimal.68 
 
Elimination/Excretion 
“In healthy volunteers (n=12) 40 – 65% of 1 mg orally administered colchicine was recovered 
unchanged in urine. Enterohepatic recirculation and biliary excretion are also postulated to play a 
role in colchicine elimination. Following multiple oral doses (0.6 mg twice daily), the mean 
elimination half-lives in young healthy volunteers (mean age 25 to 28 years of age) is 26.6 to 
31.2 hours.”68 
 
Preparation of Investigational Colchicine by Pharmaceutical Development Section or Pine 
Pharmaceuticals 
Colchicine USP powder, purchased from Spectrum Chemical, will be dispensed. Each capsule 
will contain 600 micrograms of colchicine. Matching placebo capsule will be produced that only 
contain the non-colchicine ingredients. Colchicine will be verified to be chemically pure and to 

49 
 
have acceptable stability using standard approaches by the NIH Pharmaceutical Development 
Section (before 2017), the NIH IDMS subsequently, or Pine Pharmaceuticals. 
 
I. Recruitment Strategies 
Participants will be recruited from Washington, DC and local suburbs in Maryland and Virginia. 
Recruitment efforts will aim to enroll subjects in proportion to their representation in the 
population of Maryland. Subjects of all racial, ethnic and socio-economic backgrounds will be 
invited to participate in this study.  We will contact by mail or email, adult obese subjects who 
have previously indicated interest in research and/or have already participated in protocols run 
by the Section on Growth and Obesity (SGO), to inform them of this new study and assess their 
potential interest.  These potential subjects will be provided with a telephone number and email 
address to call or write if they are interested to request more information. Participants will be 
also be recruited by means of flyers posted in the NIH Clinical Center, in other buildings at the 
NIH campus in Bethesda, MD, and in surrounding local businesses.  If necessary, further 
subjects will be recruited through newspaper advertisements or letter mailings. Advertisements 
will be submitted to the IRB before use. 
 
J. Existing data: n/a 
 
K. Description of criteria for withdrawal from study 
 
 
The types, frequency, and duration of tests and visits are outlined on the flow diagrams 
(Appendix A1-6).  Subjects will be examined at each of their visits. If unanticipated reactions are 
uncovered during the study, we will report these promptly to the IRB. The CRC has ample medical 
resources should any unanticipated event occur. Additionally, study staff will seek evidence of 
emotional or physical distress or discomfort, and will be instructed to terminate the protocol if a 
participant demonstrates distress. 
 
Study staff will emphasize to participants that they are able to withdraw from the study at 
any time, without penalty. Noncompliance with study procedures may lead to withdrawal of 
study subjects – for example, unwillingness to take the medication or inability to complete 
required testing (e.g. FSIVGTT). Other events that will result in discontinuation of treatment 
include:  
1.  Pregnancy. 

50 
 
2.  Any verified abnormalities in laboratory values or physical status that cannot be explained 
by an intercurrent unrelated illness (or by obesity itself) that are considered CTCAE grade 2 or 
greater for gastrointestinal, hematologic, clinical hemorrhage, kidney/bladder, alopecia, 
pulmonary, cardiac, neurocerebellar, allergy, flu-like symptoms, metabolic, or eye toxicities; and 
grade 3 or greater for infection, anorexia, circulatory, neurologic, dermatologic, coagulation, 
endocrine, or performance status toxicities.115  Problem-specific stopping rules have been 
defined in the CTCAE v4.03 submitted to the IRB in conjunction with this protocol.   
 
Unblinding Procedure 
In the randomized portion of the trial, all participants, Study Site staff, and pathology 
and laboratory personnel are blinded to the individual assignment of the order in which 
colchicine and placebo are administered. Participants will learn their individual assignments (i.e., 
be unblinded) when the trial is complete. In rare instances, it may be necessary to unblind a 
participant's order of assignment before completion of the trial, to a physician, to the Study Site, 
or to the participant.  
In general, however, participants should not be unblinded. Anytime a staff member 
learns the treatment assignment of a participant, the staff member is at risk of being influenced 
by that information. For example, if the unblinded participant was known to have taken the 
active agent and reported a toxicity, the staff member might make inferences about the effects of 
colchicine on that participant, and possibly on other participants as well. The management of 
future participants may be influenced by this information. Maintaining the double-blind design 
offers the best protection against this potential bias. 
In the unlikely event that a Serious and Unexpected Suspected Adverse Reaction 
(SUSAR) occurs and a physician must be unblinded to the treatment group to which an affected 
trial subject belongs, the Investigational Drug Management Section (IDMS) will be contacted by 
a study investigator (usually by the Principal Investigator). The study investigator will request 
that IDMS release the needed information and specify the individual to receive the information. 
IDMS will normally supply unblinding information only to a physician who is not a co-
investigator on the study and make clear that group assignment must not be given to study 
investigators. This procedure is intended to ensure that the identity of the investigational 
medicinal product is revealed only insofar as is necessary. As much as possible, the blinding 
should be maintained for the investigators and for anyone responsible for data analysis or 
interpretation. A subject’s treatment group should not be revealed to a study investigator except 
in a medical emergency, i.e. when this appears necessary to ensure the subject’s safety and 

51 
 
would be instrumental in further treatment decisions. If a subject’s treatment is unblinded, this 
must be documented by IDMS in their written records. In all cases, the reasons and rationale for 
unblinding will be also documented in writing and maintained in the study file. 
Unblinded data are also available to the Data Safety Monitoring Committee (DSMC). In 
the case of a SUSAR, the DSMC is informed and will thus have the opportunity to review 
subject-level data and make any changes to the experimental plan that are required. In the case of 
a SUSAR, the DSMC may elect to reveal the treatment group to which a subject belongs, request 
additional safeguards, or stop the study. The DSMC will assist investigators in deciding if a 
SUSAR requires any of the study investigators to become unblinded so that the FDA can be 
informed of subject assignment because of a SUSAR. 
 
L. Re-evaluation of Study: Not Applicable for this pilot study. 
 
M. Collection, monitoring, analysis and reporting of adverse events 
Adverse events, protocol deviations, unanticipated problems (UP), serious adverse events, 
sponsor and serious, are defined as described in NIH HRPP SOP 16 (“Reporting Requirements 
for Unanticipated Problems, Adverse Events and Protocol Deviations.”). All adverse events 
occurring during the study, including those observed by or reported to the research team, will be 
recorded. Serious unanticipated problems and serious protocol deviations will be reported to the 
IRB and CD as soon as possible but not more than 7 days after the PI first learns of the event. 
Not serious unanticipated problems will be reported to the IRB and CD as soon as possible but 
not more than 14 days after the PI first learns of the event. Not serious protocol deviations will 
be reported to the IRB as soon as possible but not more than 14 days after the PI first learns of 
the event. 
 
Deaths will be reported to the Clinical Director within 7 days after the PI first learns of 
the event. 
 
The PI and AIs will immediately report SAEs to the Sponsor according to the requirements of 21 
CFR 312.64(b) and as agreed upon with the sponsor. The PI will record nonserious AEs and 
report them to the Sponsor according to the timetable for reporting specified in the protocol (21 
CFR 312.64(b)). 
 

52 
 
a. Waiver of Reporting to the IRB of anticipated minor protocol deviations and minor 
adverse events (AE) 
The following anticipated minor deviations in the conduct of the protocol will not be 
reported to the IRB unless they occur at a rate greater than that which is anticipated to 
occur: 
Problem 
Expected Frequency 
Rescheduled appointment for protocol visit 
50% 
Unable to be contacted for follow up phone call 
20% 
Inability to obtain blood sample after 2 attempts 
25% 
Venous sample at a time point not obtained 
25% 
Bruising, fainting or lightheadedness after a blood draw 
10% 
Hemolysis or other issue preventing analysis of a blood or urine 
sample 
25% 
Stool sample not obtained or obtained on a different day 
25% 
Adequate subcutaneous adipose tissue sample not obtained 
25% 
Subject needs to make an additional visit for labwork (e.g. 
arrives too late for bloodwork, specimen result 
hemolyzed/equivocal, etc.) 
30% 
CT at time point not obtained 
15% 
Loss of investigational medications by subject 
15% 
Inadvertent loss of samples or data 
10% 
Appointment conducted >2 weeks beyond anticipated date 
10% 
Extra samples of blood drawn 
10% 
Inadvertent collection of additional data (e.g. extra survey, food 
record, stool sample, etc.) 
10% 
Becoming physically ill (e.g. abdominal pain, nausea, etc.) from 
consuming NIH CC food 
10% 
Sustaining minor bodily injury (e.g. twisted ankle) while 
participating in protocol 
5% 
Subject missed at least one dose but took at least 50% of study 
medication prescribed 
100% 
Subject took fewer than 50% of study medication prescribed 
15% 
Subject took an extra dose of study medication 
5% 
Subject forgot to return study medications at subsequent visit 
20% 

53 
 
Loss of, or forgot to bring back, MEMSCap 
20% 
MEMSCap not dispensed with open-label colchicine 
20% 
 
The following anticipated non-UP adverse events will not be reported to the IRB unless 
they occur at a rate greater than that known to occur in this population: 
i. Colchicine administration (events detailed in the FDA-approved package insert for 
colchicine): We anticipate that up to 30% of subjects will experience gastrointestinal 
side effects (e.g. diarrhea, abdominal pain, nausea, vomiting),68 which may require 
dose reductions to 0.6mg once daily. If this dose adjustment does not improve 
symptoms, it will be decided by patient and physician, on a case by case basis, 
whether to discontinue medication before the study is completed. Other anticipated 
adverse events from colchicine include: fatigue (4%), pharyngolaryngeal pain (3%), 
and headache (2%). Rare events (< 1%) may include alopecia, blood dyscrasias, 
hepatotoxicity, hypersensitivity reaction, neuromuscular toxicity (e.g. myalgia, 
myasthenia, myopathy, neuropathy, dizziness, rhabdomyolysis), and rash.68 
If the rate or severity of these events exceeds the rate or severity anticipated in the 
protocol or FDA-approved package insert, the events will be classified and reported as 
though they are Unanticipated Problems. 
 
Because we anticipate that other mild symptoms may occur in the course of 
participation in the protocol unrelated to study drug or participation, we will not 
submit Unanticipated Problems to mild signs/symptoms (CTCAE Grade ≤ 2), such as 
rhinorrhea, nasal congestion, watery eyes, cough, lightheadedness, mild wheezing, 
menstrual cramps, muscle weakness, fever blister, mild urinary tract infection, etc., 
that occur infrequently (≤ 10%).   
 
ii. OGTT: Infection (<5%), bruising (<30%), phlebitis (10%), nausea (10%), vomiting 
(10%), lightheadedness (10%).  
iii. FSIVGTT: Infection (<5%), bruising (<30%), phlebitis (10%), nausea (10%), 
vomiting (10%), hypoglycemia (10%), lightheadedness (10%), hunger (10%).  
iv. Fat Biopsy: Infection (<5%), bruising (<30%), bleeding (10%), nausea (10%), 
lightheadedness (10%), soreness (50%), local skin reaction (10%). 
v. Other: Skin reaction to EKG electrodes <15% 
 

54 
 
If the rate of these events exceeds the rate specified in the protocol, the events will be 
classified and reported as though they are Unanticipated Problems.  
 
b. Plan to monitor and report adverse events  
As outlined in Appendix A, for randomized and open-label subjects, adverse events will 
be monitored via telephone at one week, one month, and two months after the Baseline 
Visit using structured case report forms.  If any participant develops any concerning 
adverse events, the individual will be scheduled for an immediate clinic appointment 
with a trial investigator for further work up, as necessary.  Adverse events will also be 
assessed during the final visit via interim history and physical, as well as laboratory 
studies as described in Section IV C/D.  
 
Once 20 subjects have been enrolled into the randomized study, an interim analysis 
examining the rate of unanticipated problems and adverse events will be performed to 
evaluate whether they are occurring at a frequency greater than anticipated.   
 
c. DSMB 
A DSMB will be assembled for oversight of the randomized phase of this protocol by 
the Clinical Director, NICHD, with membership and scope determined by applicable 
institutional guidelines.  The DSMB will meet to evaluate all serious adverse events, and 
will at a minimum convene after at least 20 subjects have been enrolled.  The DSMB 
will examine whether there is evidence of safety and efficacy for colchicine.  The 
primary outcome measure that we propose the DSMB examine for efficacy is change in 
insulin sensitivity as determined from the FSIVGTT using the minimal model 
calculation.  For safety, the DSMB will review adverse event questionnaire data, and 
reports of side effects from all visits.  These reports will also be supplemented by a 
manual review by the investigators, who will add any other events not captured by these 
systems, and data files of laboratory values for CBC, acute care, hepatic, and mineral 
panels.  Results will be presented as an A vs. B analysis prepared by the data manager.  
The DSMB will be asked to prepare a detailed report to supply to the IRB that describes 
any adverse events discussed, and that explains the DSMB's vote to allow the study to 
continue or to end the study.  
 
N.  IND Monitoring Plan 

55 
 
1.   
Purpose 
The purpose of this Monitoring Plan is to describe the rationale and process for the collection, 
recording, and verification of data for NICHD Protocol, titled: Pilot study of the effects of 
colchicine in non-diabetic adults with metabolic syndrome. 
 
2.   
Objectives 
a. To establish a monitoring plan to ensure the protocol data are in compliance with 
Good Clinical Practice (GCP), NICHD Institutional Review Board (IRB) and 
Federal regulations.  
b. To ensure the validity, accuracy and integrity of the data 
 
 
3.   
Study Staff Responsibilities  
Jack A. Yanovski, MD, PhD (the principal investigator) is responsible for all aspects of the 
study. Some responsibilities may be delegated to the current associate investigator Andrew 
Demidowich, MD: 
 
Delegation of responsibility will be documented on a study staff Signature and Delegation of 
Responsibility Log.  
STUDY STAFF NAME 
TITLE 
*DELEGATED RESPONSIBILITIES 
Jack Yanovski, MD, PhD 
Principal-
Investigator 
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 
Andrew Demidowich, MD 
Lead Associate 
Investigator 
1, 2, 3, 4, 5, 6, 7, 8, 11, 12 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Jordan Levine, BS 
Co-investigator 
1, 6, 7, 8, 11 
 
 
 
 

56 
 
*Delegated Study Tasks:    
1. Obtain Informed 
Consent 
2. Obtain Medical History 
3. Perform Physical Exam 
4. Assess Eligibility Criteria 
5. Prescribe Study 
Drug/device 
6. CRF Completion  
7. CRF Queries 
8. Query completion 
9. Maintain Regulatory Docs  
10. Maintain IRB documents  
11. Data Monitoring 
12. Safety Monitoring  
 
 
4.   
Source Documentation and Case Report Forms 
Jack A. Yanovski, MD, PhD and Andrew Demidowich, MD are responsible for coordinating data 
collection and will review the data for accuracy and completeness within seven days of each 
subject visit.  
 
The PI (Jack A. Yanovski, MD, PhD) along with study co-investigator Andrew Demidowich, 
MD will conduct initial monitoring. Primary documents, Case Report Forms, and the 
Clinical Trials Database will also be audited per NICHD guidelines by Amarex. LLC. 
Patient consent documents, primary outcome and safety laboratory results, and diagnostic 
test results will be monitored for accuracy, correct dating, and agreement between case 
report forms and source documents. As case report forms are entered electronically into the 
NICHD Clinical Trials Database, the computer system contains logs indicating changes 
made and the circumstances leading to these changes. 
 
FDA regulatory requirements (annual reports, adverse events reporting, etc.) related to IND 
# 120722 (Colchicine) will also be monitored. The medical records of active subjects 
(defined as subjects receiving study medication) will be monitored quarterly or more 
frequently as required.  The FDA issues will be monitored at least annually. Any major 
findings will be summarized in writing and reported to the NICHD Institutional Review 
Board and DSMC, if indicated. Investigator credentials, training records, and the delegation 
of responsibility log will also be reviewed on an annual basis. 
 

57 
 
The NICHD Data Safety Monitoring Board will also conduct monitoring per NICHD 
guidelines. They will conduct A versus B analyses on data, but may also see unblinded data 
from the trial for their review of adverse events. 
 
5.   
IRB and DSMC Documentation  
All IRB documentation can be found in PTMS. The Principal Investigator, Jack A. 
Yanovski, MD, PhD, is responsible for maintaining IRB and DSMC correspondence related 
to this protocol, including records of all reviews of the study and submissions to the IRB and 
DSMC. 
 
6.   
FDA Documentation  
Jack A. Yanovski, MD, PhD is responsible for maintaining FDA correspondence, including 
forms 1571 and 1572 and other correspondence (e.g., annual reports, amendments, safety 
reports) in the electronic binder for IND # 120722 (Colchicine).  
 
Copies of all such correspondence are also maintained in PTMS as part of the NICHD protocol record. 
 
7.   
Adverse Event Procedures and Documentation  
Please refer to Section M of the protocol. 
 
8. Study Completion 
Upon completion of the study Jack A. Yanovski, MD, PhD will retain possession of the 
electronic IND binder and electronic Protocol Binder in a secure location. FDA requires 
records be retained for at least 2 years after study completion.  HHS regulations require that 
subjects’ records be maintained for at least three years after completion of a study. 
 

58 
 
Flyers, informational email and/or letters to 
previous subjects, newspaper 
advertisements 
Inclusion and exclusion criteria are 
reviewed.  If eligible, potential subjects 
scheduled for outpatient screening visit. 
Appendix A1– Study Design 
 
Event 
 
 
 
 
 
 
Procedure 
 
  
 
 
 
 
 
 
 
 
 
 
  
   
 
 
 
 
(within 4 weeks) 
 
 
 
 
 
 
(1 week) 
 
 
 
 
 
(3 weeks later) 
 
 
 
 
 
 
 
 
 
 (1 month) 
 
 
 
 
 
 (1 month) 
 
 
 
 
 
 (3 months) 
Study advertisement 
Telephone screening 
In clinic/5-SWS Day Hospital: Interim 
H&P, baseline labs, FSIVGTT, and SAT 
biopsy will take place all on the same day.  
For subjects eligible for randomization, a 
55 day supply of pills given to subject, 
randomized to either placebo or colchicine.  
In clinic/5-SWS Day Hospital: H&P, initial 
protocol review, EKG, body composition 
DEXA, screening labs, OGTT.   
Phone Call (randomized and open-label 
subjects)
Outpatient screening visit 
By investigator: to assess for adverse 
events and adherence/tolerability.  
Baseline visit 
Interim Safety Visit (randomized and open-
label subjects)
Phone Call (randomized and open-label 
subjects) 
Final visit (randomized and open-label 
subjects) 
In 5-SWS Day Hospital: Interim H&P, 
labs, FSIVGTT, body composition DEXA, 
and SAT biopsy will take place all on the 
same day. Pill bottle will be collected from 
patient and study treatment will be 
terminated. 
In clinic/5-SWS Day Hospital: Interim 
H&P, assess for adverse events and 
adherence/tolerability, check safety labs 
(e.g. CBC, acute care and hepatic panels, 
CK). 55 day supply of pills will be given to 
the subject. 
Washout visit (optional) and discharge  
(randomized and open-label subjects) 
In 5-SWSDH: Interim H&P, labs, 
FSIVGTT, DEXA, and SAT biopsy. 

59 
 
Performed at Time 0 of the 2 hour OGTT. 
Performed in Day Hospital. 
Schedule baseline visit if subject qualifies 
Page Protocol team on arrival of patient.  
Accurate height x3 and weight.  H&P 
performed. 
Protocol is reviewed by research team, and 
consent is signed formally with witness 
present.   
Performed in Day Hospital 
Performed on 7th floor, Metabolic Unit 
Appendix A2 – Screening Visit Flowsheet 
 
                     Event 
 
 
 
 
 
 
 
Procedure 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Fasting blood draw 
2 hour oral glucose tolerance test 
Discharge from NIH 
Arrive to NIH 5-SWS Day Hospital by 
8AM 
Protocol Review 
EKG 
DXA Scan 

60 
 
Performed at Time -15 min of the 
FSIVGTT 
Performed in Day Hospital. 
Page Protocol team on arrival of patient.  
Accurate height x3 and weight.  Interval 
H&P performed. 
Performed by a study investigator. 
Performed in Radiology 
Performed in Day Hospital. Stool sample 
collected in Day Hospital if subject did not 
bring sample from home. 
Appendix A3 – Baseline Visit Flowsheet 
 
                     Event 
 
 
 
 
 
 
 
Procedure 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Fasting blood draw  
FSIVGTT 
Stool sample and  
Hollingshead Questionnaire 
Arrive to NIH 5-SWS Day Hospital by 
8AM 
Subcutaneous Adipose Tissue Biopsy 
CT Scan 
Discharge from NIH 

61 
 
Page Protocol team on arrival of patient.  
Accurate height x3 and weight.  Interval 
H&P performed. 
Performed in Day Hospital. 
Appendix A4 – Interim Safety Visit Flowsheet (randomized and open-label only) 
 
                     Event 
 
 
 
 
 
 
 
Procedure 
 
 
 
 
 
 
 
 
 
 
 
Arrive to NIH 5-SWS Day Hospital by 
8AM 
Fasting blood draw 
Discharge from NIH 

62 
 
Performed by a study investigator. 
Performed at Time -15 min of the 
FSIVGTT 
Page Protocol team on arrival of patient.  
Accurate height x3 and weight.  Interval 
H&P performed. 
Performed in Day Hospital. 
Performed on 7th floor, Metabolic Unit 
Performed in Radiology 
Performed in Day Hospital. 
Appendix A5 – Final Visit Flowsheet (randomized and open-label only) 
 
                     Event 
 
 
 
 
 
 
 
Procedure 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Subcutaneous Adipose Tissue Biopsy 
Fasting blood draw  
DXA Scan 
FSIVGTT 
Arrive to NIH 5-SWS Day Hospital by 
8AM 
CT Scan 
Exit Questionnaire 
Discharge from NIH 
Stool sample  
Collected in Day Hospital if patient did not 
bring sample from home. 

63 
 
Performed by a study investigator. 
 
Performed at Time -15 min of the 
FSIVGTT 
Page Protocol team on arrival of patient.  
Accurate height x3 and weight.  Interval 
H&P performed. 
Performed in Day Hospital. 
 
Performed on 7th floor, Metabolic Unit 
 
Performed in Radiology 
 
Appendix A6 – Optional Washout Visit Flowsheet (randomized and open-label 
only) 
 
                     Event 
 
 
 
 
 
 
 
Procedure 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Subcutaneous Adipose Tissue Biopsy 
Fasting blood draw  
DXA Scan 
FSIVGTT 
Arrive to NIH 5-SWS Day Hospital by 
8AM 
CT Scan 
Discharge from NIH 
Stool sample  
Collected in Day Hospital if patient did not 
bring sample from home. 

64 
 
Appendix B1 
 
 Potentially Significant Drug Interactions with Colchicine 
 
Drug 
Noted or Anticipated Outcome 
Strong CYP3A4 Inhibitors 
 
Atazanavir 
Clarithromycin 
Cimetidine 
Cobicistat 
Darunavir/Ritonavir 
Indinavir 
Itraconazole 
Ketoconazole 
Lopinavir/Ritonavir 
Nefazodone 
Nelfinavir 
Ritonavir 
Saquinavir 
Telaprevir 
Telithromycin 
Tipranavir/Ritonavir 
Tolbutamide 
Significant increase in colchicine plasma levels; fatal colchicine toxicity 
has been reported with clarithromycin, a strong CYP3A4 inhibitor. 
Similarly, significant increase in colchicine plasma levels is anticipated 
with other strong CYP3A4 inhibitors. 
 
 
Moderate CYP3A4 Inhibitors 
 
Amprenavir 
Aprepitant 
Azithromycin 
Diltiazem 
Erythromycin 
Fluconazole 
Fosamprenavir 
Grapefruit Juice 
Verapamil 
Significant increase in colchicine plasma concentration is anticipated. 
Neuromuscular toxicity has been reported with diltiazem and verapamil 
interactions. 
 
 
P-gp Inhibitors 
Significant increase in colchicine plasma levels; fatal colchicine toxicity 
has been reported with cyclosporine, a P-gp inhibitor. Similarly, significant 
increase in colchicine plasma levels is anticipated with other P-gp 
inhibitors. 
Cyclosporine 
Ranolazine 
 
HMG-Co A Reductase Inhibitors 
 
Atorvastatin 
Fluvastatin 
Lovastatin 
Pitavastatin 
Pravastatin 
Rosuvastatin 
Simvastatin 
The addition of one drug to a stable long-term regimen of the other has 
resulted in myopathy and rhabdomyolysis (including a fatality). 
 
 
Other Lipid Lowering Drugs 
The addition of one drug to a stable long-term regimen of the other has 
resulted in myopathy and rhabdomyolysis (including a fatality). 
Fibrates 
Gemfibrozil 
 
 
Digitalis Glycosides 
 
Digoxin 
Rhabdomyolysis has been reported. 
 
 

65 
 
Other 
Dasatinib 
Fusiric Acid (systemic) 
Ivacaftor 
Mifepristone 
 
 
Combination with colchicine may increase CYP3A4 substrates. 
Other (con’t) 
Aripiprazole 
Axitinib 
Cyanocobalamin 
Saxagliptin 
 
 
Colchicine may cause a decrease in serum concentrations. 
 
 

66 
 
Appendix B2 
 
 Medications that may confound the assessment for diabetes  
 
Thiazide diuretics at a dose greater than 25mg/day 
Systemic non-cardioselective beta-blockers 
- 
Carvedilol 
- 
Labetalol 
- 
Nadolol  
- 
Penbutolol Sulfate 
- 
Pindolol  
- 
Propanolol  
- 
Timolol 
 
Niacin 
Systemic glucocorticoids 
Prescription or over-the-counter weight loss or weight gain medications/supplements 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

67 
 
Appendix C- Surgical/Invasive Procedure Verification Checklist 
 
Date:  
 
Time:  
Procedure (PER CONSENT):  
Responsible Physician(s)/LIP(s):  
Surgical Site(s) Marked: 
 
 
  
INSTRUCTIONS: Check the boxes and sign in the appropriate boxes below to indicate the steps that have been completed. Do not 
continue with the process unless you are able to verify that each step has been completed as appropriate. 
 
Step  
  
Verification Process  
 
Signature of Care Team 
Member Documenting the 
Information 
Pre-Procedure Area:  
1)  
Identification of Correct:   
Yes  
N/A  
  
  
Patient  
  
  
  
  
Procedure  
  
  
 
  
Side, site, and level (if appropriate)  
  
  
 
2)  
Surgical Site and/or Side Marked  
  
  
  
3)  
Appropriate documents confirming the procedure side, site, and level 
where appropriate are present (for example, history and physical, nursing 
assessment, and pre-anesthesia assessment)  
  
  
 
  
Accurately completed, and signed, procedure consent form  
  
  
 
  
Correct diagnostic and radiology test results(for example, radiology 
images and scans, or pathology and biopsy reports) are properly labeled 
and available  
  
  
 
  
Required blood products, implants, devices, and/or special equipment for 
the procedure are available  
  
  
 
Immediately Prior to the Start of the Surgery/Invasive Procedure:  
4)  
“Time Out” – Prior to the procedure in the operating room/procedure 
area, ALL TEAM MEMBERS participate together in a time out to 
verbally verify:  
  
Yes  
  
N/A  
  
  
Correct patient identity  
  
  
  
  
Correct side, site, and level are marked as appropriate  
  
  
 
  
Accurately completed, and signed, procedure consent form  
  
  
 
  
Agreement on the procedure to be performed  
  
  
 
  
Correct patient position  
  
  
 
  
Availability of appropriately labeled relevant images and tests  
  
  
 
  
The need for antibiotics or fluids for irrigation purposes  
  
  
 
  
Safety precautions based on patient history or medication use  
  
  
 
*COMPLETE THIS SECTION ONLY IF UNABLE TO COMPLETE A STEP IN THE VERIFICATION PROCESS 
ABOVE – EXPLAIN WHY  
Step  
Explanation  
Signature  of Care Team Member 
Documenting the Information  
Date  
 
 
 
 

68 
 
Appendix D 
3-Month Visit (Final Visit) Exit Questionnaire 
Date _____/_____/_____ 
 
Dear Study Participant, 
 
You recently completed a research study where half of the study participants took colchicine and 
half took placebo capsules for three months.  We would like to ask you some questions about your 
feelings and attitudes about the study.  With your help we hope to be able to improve our studies in 
the future.  Please answer all questions as honestly as possible. 
 
1. 
Which capsule (colchicine or placebo) do you think you were taking?  
Check one 
 
_____  colchicine 
 
_____  placebo 
 
Why do you think so? _____________________________________________________ 
 
2. 
How sure or unsure are you about which capsules you were taking? 
 
Circle one: 
 
3. 
When you agreed to join the study, do you think that you fully understood the study and 
everything you would need to do?     Check one: 
 
 
_____ yes 
 
_____ no 
 
 
 
If no, what didn’t you understand? ___________________________________________ 
 
4. 
Knowing what you know now, if you could do it all over again, would you join this study?      
Check one: 
 
 
_____  yes 
 
_____  no 
 
5. 
What would you tell a friend who was thinking of entering the study?  Check one: 
 
 
_____  join the study  
_____  don’t join the study 
1 
2 
3 
4 
5 
Very unsure 
Somewhat 
 unsure 
Undecided 
Somewhat 
sure 
Very sure 

69 
 
 
6. 
Suppose we knew for sure that taking colchicine could help you lose one pound per year, or 
at least keep you from gaining a pound or two.  Would it be worth the effort of taking the 
colchicine? 
 
 
______ yes 
 
______ no 
 
 
7. 
Do you feel your overall health has improved due to being in the study? 
 
 
______yes 
 
______no 
 
 
8. 
What effect do you think the capsules had on your body weight? 
 
 
______ no effect 
______ lost weight 
______ gained weight  
 
9. 
If you lost weight, what do you think was the reason? 
 
Check all that apply 
 
 
______ study capsules 
     ______ changed eating habits 
 
 
______ exercise 
other (please list) _______________________________________ 
 
10. 
If you tried any special diets or weight loss supplements during the three month period 
please describe (for example, Atkins, low fat, low carb, meal bars): 
 
 
________________________________________________________________________ 
 
11. 
Do you feel the study capsules improved any of the following in you? (check all that apply): 
 
 
______ joint pain 
 
______ sense of taste 
 
 
______ mobility 
 
______ energy 
 
 
If yes, please explain: ______________________________________________________ 
 
12. 
Do you think the study capsules had any side effects? 
 
 
______ yes 
 
______ no 
 
 
If yes, please list: _________________________________________________________ 
 

70 
 
________________________________________________________________________ 
 
13. 
Did you find the alarm clock useful to help remind you to take your medication?   
(check one): 
 
 
______ yes 
 
______ no 
 
 
If no, why not? __________________________________________________________ 
 
________________________________________________________________________ 
 
14.   
Why did you decide to be part of the study? 
Rank in order of importance (1 for first choice, 2 for second choice, etc.)  
Leave blank if the reason does not apply to you at all: 
 
 
______  wanted to improve my health 
 
______  make a contribution to science / help other people 
 
______  try to lose weight 
 
______  financial compensation 
 
______  opportunity for medical care at no cost 
 
______  wanted to take colchicine 
 
______  other: ___________________________________________________________ 
 
15. 
What was the worst part of the study?   
 
Rank in order of importance (1 for first choice, 2 for second choice, etc.)  
 
Leave blank if item does not apply to you: 
 
 
______  coming to NIH (traveling, security checks) 
 
______  taking the capsules 
 
______  blood draws 
 
______  FSIVGTT (3 hour IV glucose test) 
 
______  Fat biopsy 
 
______  DEXA (body fat) scan 
 
______  Collecting the stool sample 
 
______  physical exams and body measurements 
 
______  appointment times/having to take time off from work 
 
______  filling out questionnaires 
 
______  didn’t get the results I was expecting from the study capsules 
 
______  other: ___________________________________________________________ 
 

71 
 
16. 
We know that for most people it is difficult to take every capsule on time every day and you 
probably missed some doses. What do you think might have helped you to be better at 
taking the capsules?  (check all that apply): 
 
 
 
______if I only had to take one capsule a day instead of two 
 
 
______if I could have taken the two capsules together, once a day 
 
 
______if the capsules were smaller 
 
 
______if the capsules were easier to swallow 
 
 
______if it were easier to remember to take the capsules 
 
______  other: ________________________________________________________ 
 
Please tell us anything else that may have helped you to do a better job of taking your 
capsules 
 
 
  
____________________________________________________________________ 
 
 
17. 
Please write in any other comments or suggestions for the study. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
________________________________________________________________________ 
 
THANK YOU FOR BEING A PART OF THE STUDY 

72 
 
3-Month Visit (Final Visit) Exit Questionnaire for T2DM Open Label Arm 
 
Date _____/_____/_____ 
 
Dear Study Participant, 
 
You recently completed a research study where study participants took colchicine capsules for three 
months.  We would like to ask you some questions about your feelings and attitudes about the 
study.  With your help we hope to be able to improve our studies in the future.  Please answer all 
questions as honestly as possible. 
 
1. 
When you agreed to join the study, do you think that you fully understood the study and 
everything you would need to do?     Check one: 
 
 
_____ yes 
 
_____ no 
 
 
 
If no, what didn’t you understand? ___________________________________________ 
 
2. 
Knowing what you know now, if you could do it all over again, would you join this study?      
Check one: 
 
 
_____  yes 
 
_____  no 
 
3. 
What would you tell a friend who was thinking of entering the study?  Check one: 
 
 
_____  join the study  
_____  don’t join the study 
 
4. 
Suppose we knew for sure that taking colchicine could help you lose one pound per year, or 
at least keep you from gaining a pound or two.  Would it be worth the effort of taking the 
colchicine? 
 
 
______ yes 
 
______ no 
 
 
7. 
Do you feel your overall health has improved due to being in the study? 
 
 
______yes 
 
______no 
 
 
8. 
What effect do you think the capsules had on your body weight? 
 
 
______ no effect 
______ lost weight 
______ gained weight  
 

73 
 
9. 
If you lost weight, what do you think was the reason? 
 
Check all that apply 
 
 
______ study capsules 
     ______ changed eating habits 
 
 
______ exercise 
other (please list) _______________________________________ 
 
10. 
If you tried any special diets or weight loss supplements during the three month period 
please describe (for example, Atkins, low fat, low carb, meal bars): 
 
 
________________________________________________________________________ 
 
11. 
Do you feel the study capsules improved any of the following in you? (check all that apply): 
 
 
______ joint pain 
 
______ sense of taste 
 
 
______ mobility 
 
______ energy 
 
 
If yes, please explain: ______________________________________________________ 
 
12. 
Do you think the study capsules had any side effects? 
 
 
______ yes 
 
______ no 
 
 
If yes, please list: _________________________________________________________ 
 
________________________________________________________________________ 
 
13. 
Did you find the alarm clock useful to help remind you to take your medication?   
(check one): 
 
 
______ yes 
 
______ no 
 
 
If no, why not? __________________________________________________________ 
 
________________________________________________________________________ 
 
 
 
 
 

74 
 
14.   
Why did you decide to be part of the study? 
Rank in order of importance (1 for first choice, 2 for second choice, etc.)  
Leave blank if the reason does not apply to you at all: 
 
 
______  wanted to improve my health 
 
______  make a contribution to science / help other people 
 
______  try to lose weight 
 
______  financial compensation 
 
______  opportunity for medical care at no cost 
 
______  wanted to take colchicine 
 
______  other: ___________________________________________________________ 
 
15. 
What was the worst part of the study?   
 
Rank in order of importance (1 for first choice, 2 for second choice, etc.)  
 
Leave blank if item does not apply to you: 
 
 
______  coming to NIH (traveling, security checks) 
 
______  taking the capsules 
 
______  blood draws 
 
______  FSIVGTT (3 hour IV glucose test) 
 
______  Fat biopsy 
 
______  DEXA (body fat) scan 
 
______  Collecting the stool sample 
 
______  physical exams and body measurements 
 
______  appointment times/having to take time off from work 
 
______  filling out questionnaires 
 
______  didn’t get the results I was expecting from the study capsules 
 
______  other: ___________________________________________________________ 
 
16. 
We know that for most people it is difficult to take every capsule on time every day and you 
probably missed some doses. What do you think might have helped you to be better at 
taking the capsules?  (check all that apply): 
 
 
 
______if I only had to take one capsule a day instead of two 
 
 
______if I could have taken the two capsules together, once a day 
 
 
______if the capsules were smaller 
 
 
______if the capsules were easier to swallow 
 
 
______if it were easier to remember to take the capsules 
 
______  other: ________________________________________________________ 
 

75 
 
Please tell us anything else that may have helped you to do a better job of taking your 
capsules 
 
 
  
____________________________________________________________________ 
 
 
17. 
Please write in any other comments or suggestions for the study. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
________________________________________________________________________ 
 
THANK YOU FOR BEING A PART OF THE STUDY 
 

76 
 
 
Appendix E 
MEDICATION GUIDE  
Congratulations on starting your involvement with our research protocol!  Participants in the 
Randomization Arm of this study have been randomized to receive either the research medication 
(colchicine) 0.6 mg capsules twice a day or a placebo.  Neither the study subjects nor the 
investigators know which “version” of the research medication you are receiving.  Participants in 
the Open-Label Arm of the study will be taking colchicine 0.6 mg capsules twice a day 
Read this Medication Guide before you start taking the medication.  This Guide does not take the 
place of talking to your healthcare provider or the NIH research team about your medical 
conditions or treatment. You should let your healthcare provider know that you may be taking the 
research medication as part of a research protocol.  
What is the most important information I should know about the research medication?  
The research medication can cause serious side effects or death if levels of the research 
medication are too high in your body.  
• 
Taking certain medicines with the research medication can cause your level of the research 
medication to be too high, especially if you have kidney or liver problems.  
• 
Tell the NIH research team about all your medical conditions, including if you have kidney 
or liver problems. Your dose of the research medication may need to be changed.  
• 
Tell the NIH research team about all the medicines you take, including prescription and non-
prescription medicines, vitamins and herbal supplements.  
• 
Even medicines that you take for a short period of time, such as antibiotics, can interact with 
the research medication and cause serious side effects or death.  
• 
Talk to the NIH research team, your healthcare provider, and pharmacist before taking any 
new medicine. 
  
• 
Especially tell the NIH research team if you take:  
• atazanavir sulfate (Reyataz
®
)  
 
 
• clarithromycin (Biaxin
®
)  
• cyclosporine (Neoral
®
, Gengraf
®
, Sandimmune
®
)  • darunavir (Prezista
®
)  
• fosamprenavir (Lexiva
®
) with ritonavir  
 
• fosamprenavir (Lexiva
®
)  
• indinavir (Crixivan
®
)  
 
 
 
• itraconazole (Sporanox
®
)  
• ketoconazole (Nizoral
®
) 
 
 
 
• lopinavir/ritonavir (Kaletra
®
)  
• nefazodone (Serzone
®
)  
 
 
 
• nelfinavir mesylate (Viracept
®
)  
• ritonavir (Norvir
®
)   
 
 
 
• saquinavir mesylate (Invirase
®
)  
• telithromycin (Ketek
®
)  
 
 
 
• tipranavir (Aptivus
®
)  

77 
 
 
Ask the NIH research team, your healthcare provider, or pharmacist if you are not sure if you 
take any of the medicines listed above. This is not a complete list of all the medicines that can 
interact with the research medication.  
 
• 
Know the medicines you take. Keep a list of them and show it to your healthcare provider 
and pharmacist when you get a new medicine.  
• 
Keep the research medication out of the reach of children.  
 
Who should not take the research medication?  
Do not take the research medication if you have liver or kidney problems and you take certain other 
medicines. Serious side effects, including death, have been reported in these patients even when 
taken as directed. See “What is the most important information I should know about the research 
medication?”  
What should I tell the NIH research team before starting the research medication?  
See “What is the most important information I should know about the research medication?”  
Before you take the research medication tell the NIH research team about all of your medical 
conditions including if you:  
• 
have liver or kidney problems  
• 
are pregnant or plan to become pregnant. It is not known if the research medication will 
harm your unborn baby. Please tell the NIH research team immediately if you are pregnant 
or plan to become pregnant.  
• 
are breast-feeding or plan to breast-feed. The research medication passes into your breast 
milk. Due to the possible risks to the child, you are not eligible for this study if you are 
nursing. 
 
Using the research medication with certain other medicines, such as cholesterol-lowering 
medications or digoxin, can affect each other causing serious side effects.  Talk to the NIH research 
team and your healthcare provider about whether the medications you are taking might interact with 
the research medication, and what side effects to look for.  
How should I take the research medication?  
• 
Take the research medication exactly as the NIH research team instructs you to take it. If 
you are not sure about your dosing, call the NIH research team. 
• 
The medication can be taken with or without food.  
• 
If you take too much the research medication go to the nearest hospital emergency room 
right away.  
• 
If you miss a dose, then take it as soon as you remember. If it is almost time for your next 
dose, just skip the missed dose. Take the next dose at your regular time. Do not take 2 doses 
at the same time.  
 
What should I avoid while taking the research medication?  
Avoid eating grapefruit or drinking grapefruit juice while taking the research medication. It can 
increase your chances of getting serious side effects.  

78 
 
What are the possible side effects of the research medication?  
The research medication can cause serious side effects or even cause death. See “What is the 
most important information I should know about the research medication?”  
Get medical help right away, if you have:  
• 
Muscle weakness or pain  
• 
Numbness or tingling in your fingers or toes  
• 
Unusual bleeding or bruising  
• 
Increased infections  
• 
Feel weak or tired  
• 
Have significant dizziness 
• 
Pale or gray color to your lips, tongue, or palms of your hands  
• 
Severe diarrhea or vomiting  
 
The most common side effects of the research medication are abdominal pain, diarrhea, nausea 
and vomiting.  
Tell the NIH research team if you have any side effect that bothers you or that does not go away.  
These are not all of the possible side effects of the research medication. For more information, 
ask the NIH research team, your healthcare provider, or pharmacist.  
How should I store the research medication?  
• 
Store at room temperature between 68° and 77°F (20° to 25°C).  
• 
Keep tightly closed in the provided container.  
• 
Keep out of the light.  
 
Keep the research medication and all medicines out of the reach of children.  
 

79 
 
Appendix F 
(Adapted from Common Terminology Criteria for Adverse Events v4.0 (CTCAE) 
Publish Date: May 28, 2009) 
 
Grades 
 
Grade refers to the severity of the adverse event (AE). The CTCAE displays Grades 1 through 5 with unique 
clinical descriptions of severity for each AE based on this general guideline: 
 
Grade 1: Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention not 
indicated. 
 
Grade 2: Moderate; minimal, local or noninvasive intervention indicated; limiting age-appropriate 
instrumental Activities of Daily Living (ADL)*. 
 
Grade 3: Severe or medically significant but not immediately life-threatening; hospitalization or prolongation 
of hospitalization indicated; disabling; limiting self-care ADL**. 
 
Grade 4: Life-threatening consequences; urgent intervention indicated. 
 
Grade 5: Death related to AE. 
 
*Instrumental ADL refer to preparing meals, shopping for groceries or clothes, using the telephone, 
managing money, etc. 
 
**Self-care ADL refer to bathing, dressing and undressing, feeding self, using the toilet, taking medications, 
and not bedridden. 
 

80 
 
Appendix G 
 
You will be completing Self Sampling at home (or at the NIH) for up to 4 visits as part of the 
colchicine study: 
 Screening Visit 
 Medication Initiation Visit 
 3 month Final (Medication Completed) Visit 
 6 month Washout Visit (Optional) 
 
 
Supply list for Stool Collection: 
 
1. Fecocainter stool container with toilet “frame” 
2. Brown Top Stool Collection Vial (x3) 
3. Permanent Marker 
4. Clear Bag for stool collection vial (biohazard bag) 
5. Disposable gloves 
6. Stool sampling instructions 
7. Food Dietary record + instructions 
 
 
For Questions Contact: 
Jordan Levine, Research Coordinator 
 
 
[Phone Number Written Here] 
 
jordan.levine@nih.gov 
Ginika Onyekaba, Research Coordinator 
 
 
[Phone Number Written Here] 
ginikanwa.onyekaba@nih.gov 
Andrew Demidowich, MD, Lead Associate Investigator 
[Phone Number Written Here] 
andrew.demidowich@nih.gov 
  
 
 

81 
 
STOOL SPECIMEN COLLECTION: 
1. Wash your hands. 
2. Label your brown-lid container with the date and time of collection using the 
permanent marker provided. The inside of the container is sterile until opened. 
 
 
3. To minimize urine collection, first empty your bladder before performing the next 
steps. 
 
4. Place the stool-collecting frame and Fecocontainer into the toilet, securely under 
the seat.  
          
                       
 
5. Take a bowel movement in the bowl.  

82 
 
 
6. Put on gloves. 
 
7. As shown in Figure A, unscrew the cap carefully to avoid spilling fluid.  Use the 
small spoon attachment on the lid to scoop up a “pea size” sample. Place the 
shovel with the sample into the specimen container, and secure the lid tightly and 
shake vial for 5 seconds.  Try not to spill any of the liquid inside the vial (it helps 
preserve the stool).   
 
               
  
 
8. Repeat two more times for a total of three stool vials.  Uncap and work with only 
ONE vial at a time.  
9. Place all three stool vials into the biospecimen bag provided. 
 
A 
B 
C 

83 
 
10. Dispose of remaining feces into toilet and flush (optional).  Close Fecocontainer 
with the lid provided and throw away into a garbage bag. Throw the gloves away in 
the same bag. Wash Hands. 
  
11. Return stool samples during your scheduled clinic visit.  Because the vials have the 
special preservative, they do not need to be refrigerated or frozen.  However, you 
put the bag in the freezer if you wish.   

84 
 
Colchicine Study (14-CH-0119) 
Keeping Food Records 
 
 
To better understand what you usually eat and drink when you are not in the hospital, 
please write down everything you eat and drink for a typical day. 
 
When should the food records be kept?   Write down all foods and beverages you 
consume for 24 hours before you come to the Clinical Center for your appointment. This 
day should represent a fairly typical day for you.   
 
Eat normally on this day.  Don’t change your food choices, methods of food preparation, or 
where you eat just because you are recording your intake. There is no right or wrong way 
to eat for this evaluation. 
 
How and what should be recorded?  Use a new form each day.  It is best to record what 
you eat immediately after each meal and snack. 
 
You should write down: 
 
• the time you begin eating each meal and snack 
• all foods and beverages, except for plain water. 
• all condiments (such as ketchup, margarine, mayonnaise, salad dressing, sauces, 
gravy,  sugar, etc).  You do not need to record salt, pepper, herbs and spices. 
• all medicines, vitamins, minerals, and/or other supplements taken. 
 
 
For each food and beverage that you list, include an amount and a description.  To guide 
you, refer to the sample food record and the hints for recording amounts and description 
information on the next few pages.  
 
What do I do with the forms after they are filled out?   Bring them back with you to the 
NIH when you come next time.  
 
 
If you have any questions or concerns, please contact me by e-mail or by phone. 
 
 
 
 
 
Dietitian’s name:_Amber Courville______ 
Phone number: _ 301-594-8051 _________ 
E-mail: __ courvillea@nih.cc.gov _______________ 

85 
 
Hints for Completing the Amount Column 
 
 
A.  Measure foods after preparation and cooking is completed. 
 
B.  Measurements can be listed in 4 ways: 
 
 
1.  The number of items. 
 
 
 
Examples: 
saltines  
6 
 
 
 
 
white bread 2 slices 
 
 
 
 
grapes 
12 
 
 
 
 
ketchup 
2 packets 
 
 
2.  In household measures, using standard level measuring cups and spoons. 
 
 
 
Examples: 
applesauce 1/3 cup 
 
 
 
 
popcorn 
3 cups 
 
 
 
 
jelly 
 
1 ½  TB [tablespoons] 
 
 
 
 
2% milk 
1.5 CP [cup] 
 
 
 
(Do not use non-standard measures like “handful” or “serving”) 
 
 
3.  By weight or by volume, as listed on a package (or by using a kitchen scale if         
      
you have one).  
 
 
 
Examples: 
ginger ale 
12 FL OZ [fluid ounces] 
 
 
 
 
yogurt  
4.5 oz [ounces] 
 
 
 
 
Almond Joy 49 gm [grams] 
 
 
 
 
roast beef 
2 ½  oz 
 
 
 
 
pretzels 
1/3 of 6.5 oz bag 
 
 
 
 
 
 
4.  By dimensions, using a ruler. 
 
 
 
Examples: 
pancake 
5” diameter 
 
 
 
 
meatball 
1 ¼” diameter 
 
 
 
 
lasagna  
3 ½” x 4” x 1½” cube 
 
 
 
 
pizza  
1/8 of 14” diameter  [or draw it with dimensions] 
 
 
 
 
 
 
 
• Remember not all food that is served is eaten, and at other times, you may go back for seconds.  
You may need to adjust portion sizes to reflect the amount you actually ate. 
 
 

86 
 
Hints for Completing the Description Column 
 
A.  Describe foods completely. 
 
Examples: 
 
 
 
sirloin steak, fat partially trimmed 
 
 
80% lean ground beef (or 80/20 ground beef) 
 
 
chicken drumstick, skin removed before cooking 
 
 
reduced fat or 2% milk 
 
 
baked potato, skin eaten 
 
 
carrot cake with cream cheese frosting 
 
B.  Include brand names whenever possible.  Also include terms like calcium-fortified, light, and 
reduced calorie if listed on the label. 
 
Examples: 
 
 
Country Crock Light spread tub margarine  
Oreo reduced-fat cookies 
Miller Lite draft beer 
 
 
Hellmann’s fat-free cholesterol-free mayonnaise 
 
 
Dannon lowfat fruited yogurt 
 
 
sliced peaches in light syrup 
 
C.  Include information about preparation and cooking methods. 
 
Examples: 
 
 
skinless chicken breast 
floured and pan-fried in corn oil 
 
 
canned corn  
 
tub margarine and sugar added 
 
 
Campbell’s tomato soup 
made with water 
 
 
mac and cheese 
 
from mix, made with whole  milk and  
 
 
 
 
 
 
 
       Parkay stick margarine 
 
D.  For mixed dishes and recipe items, you only need to list major ingredients.  You do not need to 
write down the entire recipe. 
 
Examples: 
 
 
potato salad  
 
made with potatoes, eggs, regular mayo 
 
 
chocolate chip cookies 
homemade with real butter, walnuts added 
 
 
meatloaf 
 
 
made with 85/15 ground beef, oatmeal,   
 
 
 
 
 
      
   ketchup, egg 
 
E.  For fast food items from major chains, you only need to name the item.  No description is 
necessary, unless you “special order” an item.  Also, note if all of it wasn’t eaten. 
 
Examples: 
 
 
Biggie Fries (Wendy’s) 
 
 
 
Quarter Pounder with Cheese (McDonald’s) - didn’t eat pickles 
 
 
Meat-lovers Pan Pizza (Pizza Hut) 
 
F.  Remember to record any additions made at the table, such as margarine, sugar, ketchup, 
mustard, sauces, mayonnaise.  List them separately, and include amounts. 
 

References: 
 
1. 
Murphy NF, MacIntyre K, Stewart S, Hart CL, Hole D, McMurray JJ. Long-term 
cardiovascular consequences of obesity: 20-year follow-up of more than 15 000 middle-aged 
men and women (the Renfrew-Paisley study). European heart journal 2006;27:96-106. 
2. 
Ogden CL, Carroll MD, Curtin LR, McDowell MA, Tabak CJ, Flegal KM. Prevalence of 
overweight and obesity in the United States, 1999-2004. Jama-J Am Med Assoc 2006;295:1549-
55. 
3. 
Kuczmarski RJ, Flegal KM, Campbell SM, Johnson CL. Increasing prevalence of 
overweight among US adults. The National Health and Nutrition Examination Surveys, 1960 to 
1991. JAMA 1994;272:205-11. 
4. 
Stienstra R, Joosten LA, Koenen T, et al. The inflammasome-mediated caspase-1 
activation controls adipocyte differentiation and insulin sensitivity. Cell metabolism 
2010;12:593-605. 
5. 
Wen H, Gris D, Lei Y, et al. Fatty acid-induced NLRP3-ASC inflammasome activation 
interferes with insulin signaling. Nature immunology 2011;12:408-15. 
6. 
Visser M, Bouter LM, McQuillan GM, Wener MH, Harris TB. Elevated C-reactive 
protein levels in overweight and obese adults. JAMA 1999;282:2131-5. 
7. 
Ford ES. The metabolic syndrome and C-reactive protein, fibrinogen, and leukocyte 
count: findings from the Third National Health and Nutrition Examination Survey. 
Atherosclerosis 2003;168:351-8. 
8. 
Pradhan AD, Manson JE, Rifai N, Buring JE, Ridker PM. C-reactive protein, interleukin 
6, and risk of developing type 2 diabetes mellitus. JAMA 2001;286:327-34. 
9. 
Pradhan AD, Manson JE, Rossouw JE, et al. Inflammatory biomarkers, hormone 
replacement therapy, and incident coronary heart disease: prospective analysis from the Women's 
Health Initiative observational study. JAMA 2002;288:980-7. 
10. 
Cancello R, Henegar C, Viguerie N, et al. Reduction of macrophage infiltration and 
chemoattractant gene expression changes in white adipose tissue of morbidly obese subjects after 
surgery-induced weight loss. Diabetes 2005;54:2277-86. 
11. 
Vandanmagsar B, Youm YH, Ravussin A, et al. The NLRP3 inflammasome instigates 
obesity-induced inflammation and insulin resistance. Nature medicine 2011;17:179-88. 
12. 
den Engelsen C, Koekkoek PS, Gorter KJ, van den Donk M, Salome PL, Rutten GE. 
High-sensitivity C-reactive protein to detect metabolic syndrome in a centrally obese population: 
a cross-sectional analysis. Cardiovasc Diabetol 2012;11:25. 
13. 
Phillips CM, Perry IJ. Does inflammation determine metabolic health status in obese and 
nonobese adults? J Clin Endocrinol Metab 2013;98:E1610-9. 
14. 
Pai JK, Pischon T, Ma J, et al. Inflammatory markers and the risk of coronary heart 
disease in men and women. N Engl J Med 2004;351:2599-610. 
15. 
Danesh J, Kaptoge S, Mann AG, et al. Long-term interleukin-6 levels and subsequent risk 
of coronary heart disease: two new prospective studies and a systematic review. PLoS medicine 
2008;5:e78. 
16. 
Danesh J, Wheeler JG, Hirschfield GM, et al. C-reactive protein and other circulating 
markers of inflammation in the prediction of coronary heart disease. N Engl J Med 
2004;350:1387-97. 
17. 
Akbaraly TN, Hamer M, Ferrie JE, et al. Chronic inflammation as a determinant of future 
aging phenotypes. CMAJ 2013;185:E763-70. 

89 
 
18. 
Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW. Obesity is 
associated with macrophage accumulation in adipose tissue. Journal of Clinical Investigation 
2003;112:1796-808. 
19. 
Johnson AR, Milner JJ, Makowski L. The inflammation highway: metabolism accelerates 
inflammatory traffic in obesity. Immunological reviews 2012;249:218-38. 
20. 
Puri V, Ranjit S, Konda S, et al. Cidea is associated with lipid droplets and insulin 
sensitivity in humans. Proc Natl Acad Sci U S A 2008;105:7833-8. 
21. 
Lumeng CN, Bodzin JL, Saltiel AR. Obesity induces a phenotypic switch in adipose 
tissue macrophage polarization. J Clin Invest 2007;117:175-84. 
22. 
Clement K, Viguerie N, Poitou C, et al. Weight loss regulates inflammation-related genes 
in white adipose tissue of obese subjects. FASEB J 2004;18:1657-69. 
23. 
Lumeng CN, Deyoung SM, Bodzin JL, Saltiel AR. Increased inflammatory properties of 
adipose tissue macrophages recruited during diet-induced obesity. Diabetes 2007;56:16-23. 
24. 
Kanda H, Tateya S, Tamori Y, et al. MCP-1 contributes to macrophage infiltration into 
adipose tissue, insulin resistance, and hepatic steatosis in obesity. J Clin Invest 2006;116:1494-
505. 
25. 
Lumeng CN, Deyoung SM, Saltiel AR. Macrophages block insulin action in adipocytes 
by altering expression of signaling and glucose transport proteins. Am J Physiol Endocrinol 
Metab 2007;292:E166-74. 
26. 
Gao Z, Hwang D, Bataille F, et al. Serine phosphorylation of insulin receptor substrate 1 
by inhibitor kappa B kinase complex. J Biol Chem 2002;277:48115-21. 
27. 
Ramadan JW, Steiner SR, O'Neill CM, Nunemaker CS. The central role of calcium in the 
effects of cytokines on beta-cell function: implications for type 1 and type 2 diabetes. Cell 
calcium 2011;50:481-90. 
28. 
Dula SB, Jecmenica M, Wu R, et al. Evidence that low-grade systemic inflammation can 
induce islet dysfunction as measured by impaired calcium handling. Cell calcium 2010;48:133-
42. 
29. 
Osborn O, Brownell SE, Sanchez-Alavez M, Salomon D, Gram H, Bartfai T. Treatment 
with an Interleukin 1 beta antibody improves glycemic control in diet-induced obesity. Cytokine 
2008;44:141-8. 
30. 
Westermark P, Andersson A, Westermark GT. Islet amyloid polypeptide, islet amyloid, 
and diabetes mellitus. Physiol Rev 2011;91:795-826. 
31. 
Grant RW, Dixit VD. Mechanisms of disease: inflammasome activation and the 
development of type 2 diabetes. Frontiers in immunology 2013;4:50. 
32. 
Guarda G, Zenger M, Yazdi AS, et al. Differential expression of NLRP3 among 
hematopoietic cells. J Immunol 2011;186:2529-34. 
33. 
Barker BR, Taxman DJ, Ting JP. Cross-regulation between the IL-1beta/IL-18 
processing inflammasome and other inflammatory cytokines. Current opinion in immunology 
2011;23:591-7. 
34. 
Dinarello CA. A clinical perspective of IL-1beta as the gatekeeper of inflammation. Eur J 
Immunol 2011;41:1203-17. 
35. 
Dinarello CA, Novick D, Kim S, Kaplanski G. Interleukin-18 and IL-18 Binding Protein. 
Frontiers in immunology 2013;4:289. 
36. 
Major CD, Gao ZY, Wolf BA. Activation of the sphingomyelinase/ceramide signal 
transduction pathway in insulin-secreting beta-cells: role in cytokine-induced beta-cell death. 
Diabetes 1999;48:1372-80. 

90 
 
37. 
van Dijk SJ, Mensink M, Esser D, Feskens EJ, Muller M, Afman LA. Responses to high-
fat challenges varying in fat type in subjects with different metabolic risk phenotypes: a 
randomized trial. PLoS ONE 2012;7:e41388. 
38. 
Youm YH, Adijiang A, Vandanmagsar B, Burk D, Ravussin A, Dixit VD. Elimination of 
the NLRP3-ASC inflammasome protects against chronic obesity-induced pancreatic damage. 
Endocrinology 2011;152:4039-45. 
39. 
Stienstra R, van Diepen JA, Tack CJ, et al. Inflammasome is a central player in the 
induction of obesity and insulin resistance. Proc Natl Acad Sci U S A 2011;108:15324-9. 
40. 
Duewell P, Kono H, Rayner KJ, et al. NLRP3 inflammasomes are required for 
atherogenesis and activated by cholesterol crystals. Nature 2010;464:1357-61. 
41. 
Martinon F, Petrilli V, Mayor A, Tardivel A, Tschopp J. Gout-associated uric acid 
crystals activate the NALP3 inflammasome. Nature 2006;440:237-41. 
42. 
Feig DI, Kang DH, Johnson RJ. Uric acid and cardiovascular risk. N Engl J Med 
2008;359:1811-21. 
43. 
Butler AE, Janson J, Bonner-Weir S, Ritzel R, Rizza RA, Butler PC. Beta-cell deficit and 
increased beta-cell apoptosis in humans with type 2 diabetes. Diabetes 2003;52:102-10. 
44. 
Donath MY, Ehses JA, Maedler K, et al. Mechanisms of beta-cell death in type 2 
diabetes. Diabetes 2005;54 Suppl 2:S108-13. 
45. 
Owyang AM, Maedler K, Gross L, et al. XOMA 052, an anti-IL-1{beta} monoclonal 
antibody, improves glucose control and {beta}-cell function in the diet-induced obesity mouse 
model. Endocrinology 2010;151:2515-27. 
46. 
Sauter NS, Schulthess FT, Galasso R, Castellani LW, Maedler K. The antiinflammatory 
cytokine interleukin-1 receptor antagonist protects from high-fat diet-induced hyperglycemia. 
Endocrinology 2008;149:2208-18. 
47. 
Ofei F, Hurel S, Newkirk J, Sopwith M, Taylor R. Effects of an engineered human anti-
TNF-alpha antibody (CDP571) on insulin sensitivity and glycemic control in patients with 
NIDDM. Diabetes 1996;45:881-5. 
48. 
Klover PJ, Clementi AH, Mooney RA. Interleukin-6 depletion selectively improves 
hepatic insulin action in obesity. Endocrinology 2005;146:3417-27. 
49. 
Weisberg SP, Hunter D, Huber R, et al. CCR2 modulates inflammatory and metabolic 
effects of high-fat feeding. J Clin Invest 2006;116:115-24. 
50. 
Kim JE, Lee MH, Nam DH, et al. Celastrol, an NF-kappaB inhibitor, improves insulin 
resistance and attenuates renal injury in db/db mice. PLoS ONE 2013;8:e62068. 
51. 
Stanley TL, Zanni MV, Johnsen S, et al. TNF-alpha antagonism with etanercept 
decreases glucose and increases the proportion of high molecular weight adiponectin in obese 
subjects with features of the metabolic syndrome. J Clin Endocrinol Metab 2011;96:E146-50. 
52. 
Pradhan AD, Cook NR, Manson JE, Ridker PM, Buring JE. A randomized trial of low-
dose aspirin in the prevention of clinical type 2 diabetes in women. Diabetes Care 2009;32:3-8. 
53. 
Ridker PM, Howard CP, Walter V, et al. Effects of interleukin-1beta inhibition with 
canakinumab on hemoglobin A1c, lipids, C-reactive protein, interleukin-6, and fibrinogen: a 
phase IIb randomized, placebo-controlled trial. Circulation 2012;126:2739-48. 
54. 
Rissanen A, Howard CP, Botha J, Thuren T. Effect of anti-IL-1beta antibody 
(canakinumab) on insulin secretion rates in impaired glucose tolerance or type 2 diabetes: results 
of a randomized, placebo-controlled trial. Diabetes Obes Metab 2012. 
55. 
Cavelti-Weder C, Babians-Brunner A, Keller C, et al. Effects of gevokizumab on 
glycemia and inflammatory markers in type 2 diabetes. Diabetes Care 2012;35:1654-62. 
56. 
Larsen CM, Faulenbach M, Vaag A, et al. Interleukin-1-receptor antagonist in type 2 
diabetes mellitus. N Engl J Med 2007;356:1517-26. 

91 
 
57. 
van Asseldonk EJ, Stienstra R, Koenen TB, Joosten LA, Netea MG, Tack CJ. Treatment 
with Anakinra improves disposition index but not insulin sensitivity in nondiabetic subjects with 
the metabolic syndrome: a randomized, double-blind, placebo-controlled study. J Clin 
Endocrinol Metab 2011;96:2119-26. 
58. 
Yang LP. Oral colchicine (Colcrys): in the treatment and prophylaxis of gout. Drugs 
2010;70:1603-13. 
59. 
Terkeltaub RA. Colchicine update: 2008. Seminars in arthritis and rheumatism 
2009;38:411-9. 
60. 
Bhattacharyya B, Panda D, Gupta S, Banerjee M. Anti-mitotic activity of colchicine and 
the structural basis for its interaction with tubulin. Medicinal research reviews 2008;28:155-83. 
61. 
Nuki G. Colchicine: its mechanism of action and efficacy in crystal-induced 
inflammation. Current rheumatology reports 2008;10:218-27. 
62. 
Sari I, Yuksel A, Kozaci D, et al. The effect of regular colchicine treatment on 
biomarkers related with vascular injury in newly diagnosed patients with familial Mediterranean 
fever. Inflammation 2012;35:1191-7. 
63. 
Ding AH, Porteu F, Sanchez E, Nathan CF. Downregulation of tumor necrosis factor 
receptors on macrophages and endothelial cells by microtubule depolymerizing agents. J Exp 
Med 1990;171:715-27. 
64. 
Chia EW, Grainger R, Harper JL. Colchicine suppresses neutrophil superoxide 
production in a murine model of gouty arthritis: a rationale for use of low-dose colchicine. 
British journal of pharmacology 2008;153:1288-95. 
65. 
Li Z, Davis GS, Mohr C, Nain M, Gemsa D. Inhibition of LPS-induced tumor necrosis 
factor-alpha production by colchicine and other microtubule disrupting drugs. Immunobiology 
1996;195:624-39. 
66. 
Ben-Chetrit E, Bergmann S, Sood R. Mechanism of the anti-inflammatory effect of 
colchicine in rheumatic diseases: a possible new outlook through microarray analysis. 
Rheumatology (Oxford) 2006;45:274-82. 
67. 
Misawa T, Takahama M, Kozaki T, et al. Microtubule-driven spatial arrangement of 
mitochondria promotes activation of the NLRP3 inflammasome. Nat Immunol 2013;14:454-60. 
68. 
Colcrys Prescribing Information. In: Takeda Pharmaceuticals America I, ed. Deerfield, 
IL; 2012. 
69. 
Miyachi Y, Taniguchi S, Ozaki M, Horio T. Colchicine in the treatment of the cutaneous 
manifestations of Behcet's disease. The British journal of dermatology 1981;104:67-9. 
70. 
Suehisa S, Tagami H, Inoue F, Matsumoto K, Yoshikuni K. Colchicine in the treatment 
of acute febrile neutrophilic dermatosis (Sweet's syndrome). The British journal of dermatology 
1983;108:99-101. 
71. 
Hazen PG, Michel B. Management of necrotizing vasculitis with colchicine. 
Improvement in patients with cutaneous lesions and Behcet's syndrome. Arch Dermatol 
1979;115:1303-6. 
72. 
Nidorf M, Thompson PL. Effect of colchicine (0.5 mg twice daily) on high-sensitivity C-
reactive protein independent of aspirin and atorvastatin in patients with stable coronary artery 
disease. Am J Cardiol 2007;99:805-7. 
73. 
Raju NC, Yi Q, Nidorf M, Fagel ND, Hiralal R, Eikelboom JW. Effect of colchicine 
compared with placebo on high sensitivity C-reactive protein in patients with acute coronary 
syndrome or acute stroke: a pilot randomized controlled trial. Journal of thrombosis and 
thrombolysis 2012;33:88-94. 

92 
 
74. 
Deftereos S, Giannopoulos G, Kossyvakis C, et al. Colchicine for prevention of early 
atrial fibrillation recurrence after pulmonary vein isolation: a randomized controlled study. J Am 
Coll Cardiol 2012;60:1790-6. 
75. 
Imazio M, Brucato A, Cemin R, et al. A randomized trial of colchicine for acute 
pericarditis. N Engl J Med 2013;369:1522-8. 
76. 
Imazio M, Brucato A, Cemin R, et al. Colchicine for recurrent pericarditis (CORP): a 
randomized trial. Ann Intern Med 2011;155:409-14. 
77. 
Millaire A, de Groote P, Decoulx E, Goullard L, Ducloux G. Treatment of recurrent 
pericarditis with colchicine. European heart journal 1994;15:120-4. 
78. 
Finkelstein Y, Shemesh J, Mahlab K, et al. Colchicine for the prevention of 
postpericardiotomy syndrome. Herz 2002;27:791-4. 
79. 
Imazio M, Trinchero R, Brucato A, et al. COlchicine for the Prevention of the Post-
pericardiotomy Syndrome (COPPS): a multicentre, randomized, double-blind, placebo-
controlled trial. European heart journal 2010;31:2749-54. 
80. 
Nidorf SM, Eikelboom JW, Budgeon CA, Thompson PL. Low-dose colchicine for 
secondary prevention of cardiovascular disease. J Am Coll Cardiol 2013;61:404-10. 
81. 
Deftereos S, Giannopoulos G, Raisakis K, et al. Colchicine treatment for the prevention 
of bare-metal stent restenosis in diabetic patients. Journal of the American College of Cardiology 
2013;61:1679-85. 
82. 
Ertel NH, Akgun S. Inhibition of glucose-induced insulin secretion by intravenous 
colchicine in man. Metabolism 1979;28:1255-60. 
83. 
Giugliano D, Cerciello T, Passariello N, et al. Colchicine and insulin secretion in man. 
Diabetes 1981;30:1008-12. 
84. 
Burstein R, Seidman DS, Zemer D, et al. Chronic colchicine treatment does not impair 
glucose tolerance in familial Mediterranean fever patients. Eur J Clin Pharmacol 1997;52:27-30. 
85. 
Schlesinger N, Schumacher R, Catton M, Maxwell L. Colchicine for acute gout. 
Cochrane Database Syst Rev 2006:CD006190. 
86. 
Karnieli E, Zarnowski MJ, Hissin PJ, Simpson IA, Salans LB, Cushman SW. Insulin-
stimulated translocation of glucose transport systems in the isolated rat adipose cell. Time 
course, reversal, insulin concentration dependency, and relationship to glucose transport activity. 
The Journal of biological chemistry 1981;256:4772-7. 
87. 
Lizunov VA, Matsumoto H, Zimmerberg J, Cushman SW, Frolov VA. Insulin stimulates 
the halting, tethering, and fusion of mobile GLUT4 vesicles in rat adipose cells. The Journal of 
cell biology 2005;169:481-9. 
88. 
Jimenez-Gomez Y, Mattison JA, Pearson KJ, et al. Resveratrol improves adipose insulin 
signaling and reduces the inflammatory response in adipose tissue of rhesus monkeys on high-
fat, high-sugar diet. Cell metabolism 2013;18:533-45. 
89. 
Ranganathan S, Ciaraldi TP, Henry RR, Mudaliar S, Kern PA. Lack of effect of leptin on 
glucose transport, lipoprotein lipase, and insulin action in adipose and muscle cells. 
Endocrinology 1998;139:2509-13. 
90. 
Vazquez-Prieto MA, Bettaieb A, Haj FG, Fraga CG, Oteiza PI. (-)-Epicatechin prevents 
TNFalpha-induced activation of signaling cascades involved in inflammation and insulin 
sensitivity in 3T3-L1 adipocytes. Archives of biochemistry and biophysics 2012;527:113-8. 
91. 
Handa M, Vanegas S, Maddux BA, et al. XOMA 052, an anti-IL-1beta monoclonal 
antibody, prevents IL-1beta-mediated insulin resistance in 3T3-L1 adipocytes. Obesity (Silver 
Spring) 2013;21:306-9. 
92. 
Lizunov VA, Stenkula KG, Blank PS, et al. Human adipose cells in vitro are either 
refractory or responsive to insulin, reflecting host metabolic state. PloS one 2015;10:e0119291. 

93 
 
93. 
Grundy SM, Cleeman JI, Daniels SR, et al. Diagnosis and management of the metabolic 
syndrome: an American Heart Association/National Heart, Lung, and Blood Institute Scientific 
Statement. Circulation 2005;112:2735-52. 
94. 
Ascaso JF, Pardo S, Real JT, Lorente RI, Priego A, Carmena R. Diagnosing insulin 
resistance by simple quantitative methods in subjects with normal glucose metabolism. Diabetes 
Care 2003;26:3320-5. 
95. 
Standards of medical care in diabetes--2014. Diabetes care 2014;37 Suppl 1:S14-80. 
96. 
Herman WH, Ma Y, Uwaifo G, et al. Differences in A1C by race and ethnicity among 
patients with impaired glucose tolerance in the Diabetes Prevention Program. Diabetes care 
2007;30:2453-7. 
97. 
Herman WH, Dungan KM, Wolffenbuttel BH, et al. Racial and ethnic differences in 
mean plasma glucose, hemoglobin A1c, and 1,5-anhydroglucitol in over 2000 patients with type 
2 diabetes. The Journal of clinical endocrinology and metabolism 2009;94:1689-94. 
98. 
Davidson MB, Schriger DL. Effect of age and race/ethnicity on HbA1c levels in people 
without known diabetes mellitus: implications for the diagnosis of diabetes. Diabetes research 
and clinical practice 2010;87:415-21. 
99. 
World Health Organization multicenter study on menstrual and ovulatory patterns in 
adolescent girls. II. Longitudinal study of menstrual patterns in the early postmenarcheal period, 
duration of bleeding episodes and menstrual cycles. World Health Organization Task Force on 
Adolescent Reproductive Health. Journal of adolescent health care : official publication of the 
Society for Adolescent Medicine 1986;7:236-44. 
100. 
Hollingshead AB. Hollingshead two factor index of social position (1957). In: Miller DC, 
ed. Handbook of research design and social measurement. 5th edition ed. Newbury Park, CA: 
Sage Publications; 1991:351-9. 
101. 
O'Connell MB, Wong MO, Bannick-Mohrland SD, Dwinell AM. Accuracy of 2- and 8-
hour urine collections for measuring creatinine clearance in the hospitalized elderly. 
Pharmacotherapy 1993;13:135-42. 
102. 
Markantonis SL, Agathokleous-Kioupaki E. Can two-, four- or eight-hour urine 
collections after voluntary voiding be used instead of twenty-four-hour collections for the 
estimation of creatinine clearance in healthy subjects? Pharmacy world & science : PWS 
1998;20:258-63. 
103. 
Bergman RN. Lilly lecture 1989. Toward physiological understanding of glucose 
tolerance. Minimal-model approach. Diabetes 1989;38:1512-27. 
104. 
Welch S, Gebhart SS, Bergman RN, Phillips LS. Minimal model analysis of intravenous 
glucose tolerance test-derived insulin sensitivity in diabetic subjects. J Clin Endocrinol Metab 
1990;71:1508-18. 
105. 
Jackness C, Karmally W, Febres G, et al. Very low-calorie diet mimics the early 
beneficial effect of Roux-en-Y gastric bypass on insulin sensitivity and beta-cell Function in type 
2 diabetic patients. Diabetes 2013;62:3027-32. 
106. 
Anderson RL, Hamman RF, Savage PJ, et al. Exploration of simple insulin sensitivity 
measures derived from frequently sampled intravenous glucose tolerance (FSIGT) tests. The 
Insulin Resistance Atherosclerosis Study. Am J Epidemiol 1995;142:724-32. 
107. 
Saad MF, Anderson RL, Laws A, et al. A comparison between the minimal model and 
the glucose clamp in the assessment of insulin sensitivity across the spectrum of glucose 
tolerance. Insulin Resistance Atherosclerosis Study. Diabetes 1994;43:1114-21. 
108. 
Yang YJ, Youn JH, Bergman RN. Modified protocols improve insulin sensitivity 
estimation using the minimal model. Am J Physiol 1987;253:E595-602. 

94 
 
109. 
Korytkowski MT, Berga SL, Horwitz MJ. Comparison of the minimal model and the 
hyperglycemic clamp for measuring insulin sensitivity and acute insulin response to glucose. 
Metabolism 1995;44:1121-5. 
110. 
Coates PA, Luzio SD, Brunel P, Owens DR. Comparison of estimates of insulin 
sensitivity from minimal model analysis of the insulin-modified frequently sampled intravenous 
glucose tolerance test and the isoglycemic hyperinsulinemic clamp in subjects with NIDDM. 
Diabetes 1995;44:631-5. 
111. 
Davidson LE, Kuk JL, Church TS, Ross R. Protocol for measurement of liver fat by 
computed tomography. Journal of applied physiology 2006;100:864-8. 
112. 
Speliotes EK, Massaro JM, Hoffmann U, et al. Liver fat is reproducibly measured using 
computed tomography in the Framingham Heart Study. Journal of gastroenterology and 
hepatology 2008;23:894-9. 
113. 
Fujimoto WY, Jablonski KA, Bray GA, et al. Body size and shape changes and the risk 
of diabetes in the diabetes prevention program. Diabetes 2007;56:1680-5. 
114. 
Common Terminology Criteria for Adverse Events (CTCAE) v4.03. In: National Cancer 
Institute, National Institutes Health, ed. Bethesda, MD: U.S. Department of Health and Human 
Services; 2010. 
115. 
Perry M. In: Williams, Wilkins, eds. The Chemotherapy Sourcebook. Baltimore; 1992 
1133-40. 
116. 
Essah PA, Arrowood JA, Cheang KI, Adawadkar SS, Stovall DW, Nestler JE. Effect of 
combined metformin and oral contraceptive therapy on metabolic factors and endothelial 
function in overweight and obese women with polycystic ovary syndrome. Fertility and sterility 
2011;96:501-4 e2. 
117. 
Mitri J, Dawson-Hughes B, Hu FB, Pittas AG. Effects of vitamin D and calcium 
supplementation on pancreatic beta cell function, insulin sensitivity, and glycemia in adults at 
high risk of diabetes: the Calcium and Vitamin D for Diabetes Mellitus (CaDDM) randomized 
controlled trial. Am J Clin Nutr 2011;94:486-94. 
118. 
O'Keefe JH, Jr., McCallister BD, Bateman TM, Kuhnlein DL, Ligon RW, Hartzler GO. 
Ineffectiveness of colchicine for the prevention of restenosis after coronary angioplasty. J Am 
Coll Cardiol 1992;19:1597-600. 
119. 
Chappey ON, Niel E, Wautier JL, et al. Colchicine disposition in human leukocytes after 
single and multiple oral administration. Clin Pharmacol Ther 1993;54:360-7.