A Pilot Study of Using Statins in Patients With Acute Venous Thromboembolism (VTE)
Phase: EARLY_PHASE1
Conditions: Venous Thromboembolism
Sponsor: Ohio State University

 
 
 
 
 
 
 
 
 
 
 
 
 
A pilot study of using statins in patients with acute venous 
thromboembolism 
 
 
Coordinating Center:  
 
The Ohio State University 
 
 
 
 
Principal Investigator:  
 
Tzu-Fei Wang 
 
A340 Starling Loving Hall 
 
320 W 10th Avenue 
 
Phone: 614-293-2887 
 
Fax: 614-293-7529 
 
Email: Tzu-Fei.Wang@osumc.edu 
 
 
Study drug(s):                                   Atorvastatin 
  
 
Clinical Trials.gov #:                        NCT02331095 
 
 
 
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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Schema 
 
 
 
1.0 
Objectives 
 
This will be a single-center, open-labelled, pilot study.  This study has the following objectives:  
1.1     Primary Objective 
 
1.1.1  To determine the  reduction  of  thrombin  peak  concentration  and/or endogenous 
thrombin potential measured by Thrombin Generation Assay (TGA) at 3 months in the 
“anticoagulation +atorvastatin” arm as compared to the “anticoagulation” arm. 
 
 
 1.2      Secondary Objectives 
 
1.2.1  To determine the composite rate of recurrent VTE and VTE related mortality in patients 
in the statin arm as compared to no statin arm. 
 
1.2.2  To determine the rate of arterial thrombotic events in patients in the statin arm as 
compared to no statin arm. 
 
1.2.3  To determine the rate of major, non-major, and all hemorrhages defined by the ISTH 
(International Society on Thrombosis and Haemostasis) criteria (see section 7.3) in 
patients in the statin arm as compared to no statin arm. 
 
Atorvastatin 40 mg 
Atorvastatin  

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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1.2.4  To evaluate the changes in proposed hemostatic, inflammatory, and lipidomic 
biomarkers at 3 and 9 months compared to upon enrollment in patients in both arms.  
 
1.2.5  To determine the rate of residual vein obstruction by Doppler ultrasound at 3 months in 
patients in the statin arm as compared to no statin arm. 
 
1.2.6  To determine the rate of clinical post-thrombotic syndrome (PTS), as objectively 
evaluated with Villalta scoring system in patients in the statin arm as compared to no 
statin arm. 
 
 
2.0      Background 
 
Venous thromboembolism (VTE) refers to the formation of pathological blood clots in 
extremity veins (deep vein thrombosis, DVT) and/or pulmonary arteries (pulmonary 
embolism, PE). VTE is common, with an estimated incidence of 1-2 per 1000 in the 
general population, and increases to over 1% in the elderly.1 VTE is a considerable public 
health threat. Approximately  300,000-600,000  patients  develop  VTE  in  the  United  
States  each  year, accounting for 60,000-100,000 deaths. According to the Healthcare Cost 
and Utilization Project, about 350,000 hospital admissions were due to VTE, costing over 
three billion dollars in 2011 alone. In addition, approximately one third of patients die within 
one year of their initial VTE, and one third of survivors suffer from long-term morbidity. 
 
Anticoagulation is the current standard o f  care for acute VTE. However, even 
after a course of anticoagulation, 30% patients have VTE recurrence in 10 years after an 
unprovoked VTE,2 and 25-50% patients will develop post-thrombotic syndrome (PTS), 
causing long-term morbidity. 3,4 Therefore, there is significant room for improvement. To 
accomplish this, it is crucial to have a better understanding of the pathophysiology of VTE 
and its complications. Although some important biomarkers in VTE were identified, 12-14 
the detailed hemostatic, inflammatory, and lipidomic biomarker profiles in patients after an 
acute VTE and their evolution and interplay in relation to treatments are unclear. A better 
understanding can help to identify promising targets to develop novel therapy. 
 
The first aim of the study is to investigate important biomarkers in acute VTE as below: 
 
1)  Hemostatic profile: 
In acute VTE, patients’ pro-coagulation, anti-coagulation, and fibrinolytic systems are 
highly activated. The routine clinical coagulation assays such as prothrombin time and 
activated partial thromboplastin time terminate at initial clot formation, and are inadequate for 
comprehensive assessment of the coagulation system. In contrast, thrombin generation assays 
are increasingly used to assess the global hemostatic status. High thrombin generation potential 
or high thrombin peak concentration was correlated with increased risk of first and 
recurrent VTE.15-17 However, the thrombin generation profiles in acute VTE and their changes 
with anticoagulation have not been solidified. Therefore, thrombin generation will be used as 
our primary biomarker of interest. Secondly, a product of fibrinolysis, D-dimer, indicates 
ongoing fibrin formation and degradation. Elevated D-dimer after the completion of 
anticoagulation has been shown to predict increased risk of recurrent VTE.18,19 Therefore, 
we will also follow D- dimer during and after acute VTE. Thirdly, tissue factor (TF) and 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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tissue factor pathway inhibitor (TFPI) play essential roles in the initiation and modulating of 
blood coagulation cascade and are of interest in the process of venous thrombosis. We 
therefore plan to study these biomarkers in our patient population as well.36  
 
2)  Inflammatory profiles: 
Inflammation and coagulation are closely interwined.20 Inflammatory diseases are 
associated with increased risk of VTE,21 and thrombosis is associated with release of 
inflammatory cytokines. 22However, their changes in relates to anticoagulation and statins are 
unclear, and therefore selected inflammatory biomarkers including IL-6, IL-8, tumor necrosis 
factor (TNF)-α, high sensitivity C-reactive protein (hs-CRP) will be evaluated. 
 
 
3)  Lipidomic profiles: 
Serum phospholipids play crucial roles in inflammation and serve as the substrate 
for phospholipase A2 (PLA2 ), which subsequently liberates arachidonic acid for 
cyclooxygenase (COX) and lipoxygenase (LOX) to generate eicosanoids.23In addition, these 
bioactive phospholipids play a major role in inflammation and repair.24 However, the 
quantification of these bioactive lipids and the process of their modulation of inflammation 
during acute VTE are unclear, and this study intends to clarify it. Lipid biomarkers of 
inflammation that will be analyzed in the study include free fatty acids and eicosanoids. 
 
Statins are effective in the prevention of arterial thrombosis.25 Recently, arterial and 
venous thrombosis are shown to share common pathophysiological mechanisms,26 and 
effective therapies for arterial thrombosis could provide benefits in VTE. Several observational 
studies and the JUPITER trial, a large, randomized, placebo-controlled study, demonstrates 
that statins significantly reduce the risk of first VTE by 40-50%.7, 27-30 Additionally, as few as 3 
days of atorvastatin  increase  plasma  fibrin  clot  permeability  and  susceptibility  to  
lysis.31These promising results, as well as their safety profiles, make statins an attractive 
potential addition to the standard anticoagulation for treating acute VTE, in an effort to reduce 
long-term morbidity. 
Our second aim is to determine the actions of statins on the proposed biomarkers in 
acute VTE because: First, the effects of statins on thrombin generation in patients with acute 
VTE have not been studied. A study in patients with atrial fibrillation on warfarin showed a 
40% reduction in endogenous thrombin potential with only three months of intensive 
cholesterol-lowering treatment including statins.8 Similar effects could be seen in patients with 
acute VTE. In addition, previous studies evaluating the effects of statins on the reduction of 
D-dimer or inflammatory cytokines revealed promising results but were not focused on 
patients with acute VTE.32,33 Therefore, this study will generate important information for 
acute VTE patients. 
 
3.0      Patient Selection 
 
 
3.1      Inclusion of Women and Minorities 
 
Entry to this study is open to both men and women ages 18 and older, of all racial and 
ethnic subgroups. This study will be conducted in accordance with the International 
Conference on Harmonisation and Helsinki Guidelines. 
 
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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3.2      Inclusion Criteria 
 
 
3.2.1 At least 18 years old 
 
3.2.2    A diagnosis of proximal DVT (proximal to and including popliteal vein), with or 
without PE, confirmed by objective imaging studies, such as Doppler ultrasound, 
venograms (for DVT) and/or computer tomography, angiograms, ventilation-perfusion 
scan (for PE) 
 
 
3.2.3 Treated with warfarin or rivaroxaban as anticoagulation (short-term bridging with 
heparin or lovenox, if needed is allowed) 
 
3.3      Exclusion Criteria 
 
 
3.3.1   Thrombolysis within 6 weeks prior to enrollment 
 
3.3.2    Patients with statin use within 6 weeks of enrollment  
 
3.3.3 Patients with known allergy or intolerance to statins or statins are contraindicated for any 
other reasons 
 
3.3.4 Patients with baseline aspartate aminotransferase (AST), alanine aminotransferase (ALT), 
or total bilirubin ≥2.0 x ULN (upper limit of normal) 
 
3.3.5    Pregnant or breastfeeding females are excluded  
 
3.3.6    Any  malignancy  diagnosed  within  the  preceding  2  years,  except  for  squamous  
cell carcinoma or basal cell carcinoma of skin treated with local resection only, or 
carcinoma in situ of the cervix 
 
3.3.7 Incarcerated patients are excluded from the study due to the inherent difficulties in 
maintaining close follow-up for study purposes in patients who are incarcerated.   
 
3.4      Enrollment/registration of patients 
 
3.4.1 Patients must give written informed consent prior to enrollment on this study.  Informed 
consent may be obtained by the principal investigator, co-investigators, or one of their 
designees.  
 
4.0      Randomization of Patients 
 
 
 
 
Patients will be randomized at the time of registration/enrollment on this clinical trial via 
contact with the study nurse/coordinator at the time of the initial registration of the 
subject.  The study nurse/coordinator will conduct the randomization on the REDCap 
Data Capture system and relay this information to the treating/enrolling physician so that 
the appropriate medication (atorvastatin) can be started.   
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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5.0      Treatment plan: atorvastatin vs placebo 
 
 
After randomization, patients on the statin arm will start the assigned study drug 
(atorvastatin) for a total of 9 months, in addition to the standard of care (anticoagulation). 
Atorvastatin will be prescribed through patients’ pharmacy of choice. It will be taken 
once a day in a tablet form. Pill count and patient self-report compliance diary will be 
reviewed at each visit. All the attempts will be made to continue atorvastatin for entire 9 
months, however, if a patient needs to stop atorvastatin prior to the intended period for 
any reasons (for example, side effects), it will be documented, but they are allowed to 
remain in the study for blood tests and clinical follow up. At the end of the study, the 
decision of whether patients should remain on atorvastatin or not will be left to the 
treating physicians.   
 
6.0       Study calendar 
 
 
Initial visit 
3 month1 
9 month1 
Singed informed 
consent 
x 
 
 
Medical/treatment 
history 
x 
x 
x 
Physical examination 
x 
x 
x 
Vital signs 
x 
x 
x 
CBC with diff2 
x 
x 
x 
Chemistry3, including 
liver function tests 
x 
x 
x 
Fasting lipid profiles 
x 
x 
x 
hs-CRP 
x 
x 
x 
D-dimer 
x 
x 
x 
Urine β-hCG4 
x 
 
 
Thrombin generation 
assay 
x 
x 
x 
IL-6, IL-8, TNF-α 
x 
x 
x 
Lipidomic profiles5 
x 
x 
x 
Doppler ultrasound of 
bilateral lower 
extremities 
 
x 
 
Prescription of statins6 x 
x 
 
1All scheduled visits are allowed a window of +/- 21 days  
2CBC includes: white blood cells with differential, hemoglobin, red blood cells, and platelets  
3Chemistry and liver function tests include: sodium, potassium, carbon dioxide, glucose, BUN, creatinine, calcium, 
total protein, albumin, bilirubin, alkaline phosphatase, AST and ALT  
4Required only in women of child-bearing potential 
5Lipidomic profiles includes free fatty acids, lipoprotein-associated phospholipase A2, pro-inflammatory 
eicosanoids 
6Prescription of statins only in the statin arm 
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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7.0      Outcome Criteria 
 
7.1      Recurrent venous thromboembolism 
 
7.1.1   Recurrent pulmonary embolism will be diagnosed when: 
  
a) New filling defect(s) is/are seen in CT angiogram of chest or pulmonary 
angiography 
b) A new high probability on ventilation-perfusion lung scan. 
 
7.1.2   Recurrent deep vein thrombosis will be diagnosed when: 
  
         
a)    New filling defect(s) is/are seen in venography or new uncompressible segments 
seen on vascular Doppler ultrasound in a previously uninvolved limb 
          
b)    In the limb with a previous VTE, new filling defect(s) in venography or new 
uncompressible segments in vascular Doppler ultrasound that is clearly extending 
from the prior thrombosis or involving a new venous segment 
 
7.2      Arterial thromboembolism 
 
7.2.1 Myocardial infarction: 
 
          Myocardial infarction will be diagnosed if at least two of the following criteria are met: a 
history of typical ischemic pain lasting for at least 20 minutes and unresponsive to 
sublingual nitrates (if given), new elevation of cardiac enzymes (creatinine kinase or its 
MB fraction) to more than twice the ULN or troponin value, new EKG changes including 
new persistent ST/T changes, new bundle branch block or new Q waves in at least two 
consecutive leads.  
 
7.2.2   Cerebral vascular accidents: 
 
          Cerebral vascular accidents (CVA) is a clinical syndrome of rapid development of focal or 
global loss of brain function, lasting more than 24 hours, thought to be vascular in origin, 
and/or leading to neurological damages or early death. It is expected that CT or MRI scans 
will be performed to confirm the diagnosis in most cases.  
 
7.3      Bleeding 
 
 
7.3.1 Major bleeding will be defined as: 
 
                      a) Bleeding into critical organs such as central nervous system, retroperitonium 
                      b) Bleeding requiring emergent intervention or surgery for hemostasis 
                      c) Bleeding requiring at least 2 units of packed red blood cell transfusion 
                      d) Bleeding leading to at least 2 points drop in hemoglobin from baseline 
                      e) Bleeding requiring hospital admission for intervention 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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7.3.2 Non-major bleeding is defined as any other bleeding reported by patients but not 
otherwise meeting the above listed criteria for major bleeding 
 
8.0      Treatment after Recurrences of VTE 
 
 
Patients suffering a recurrent VTE on either arm will stop study drugs and but will 
continue to be followed on the study. The management of recurrent VTE will be left to 
the discretion of the treating physician.  
9.0      Correlative Studies 
9.1       Laboratory Studies at Enrollment  
9.1.1 The initial laboratory tests will be obtained at the time of enrollment on this study. Blood 
will be collected after overnight fasting for at least 9 hours. Laboratory tests will include: 
 
 CBC (complete blood count) 
 chemistry panel (Na, K, Cl, CO2, Cr, BUN, Ca, and glucose),  
 liver function tests (AST, ALT, ALP, total and direct bilirubin) 
 D-dimer 
 high-sensitivity C-reactive protein (hs-CRP) 
 fasting lipid profiles (LDL, HLD, TG, total cholesterol 
 β-hCG is required only in women of child-bearing potential 
             
 
Research labs: 
 thrombin generation assay 
 IL-6, IL-8, tumor necrosis factor (TNF)-α 
 lipidomic profiles including free fatty acids, lipoprotein-associated phospholipase A2,  
pro-inflammatory eicosanoids 
 plasma levels and RNA expression of tissue factor (TF), tissue factor pathway 
inhibitor (TFPI), and other clotting factors  
9.2    Subsequent Follow-up and Correlative Laboratory Studies 
 
9.2.1 All enrolled patients will be seen on enrollment and at month 3 and 9.   
 
9.2.2 The following laboratory studies will be obtained at each follow-up visit: 
 
 CBC (complete blood count), platelet count  
 chemistry panel (Na, K, Cl, CO2, Cr, BUN, Ca, and glucose),  
 liver function tests (AST, ALT, ALP, total and direct bilirubin) 
 d-dimer 
 high-sensitivity C-reactive protein (hs-CRP) 
 fasting lipid profiles (LDL, HLD, TG, total cholesterol) 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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Research labs:  
 thrombin generation assay 
 IL-6, IL-8, tumor necrosis factor (TNF)-α 
 lipidomic profiles including free fatty acids, lipoprotein-associated phospholipase A2, 
pro-inflammatory eicosanoids 
 plasma levels and RNA expression of tissue factor (TF), tissue factor pathway 
inhibitor (TFPI), and other clotting factors  
 
9.2.3 A follow up Doppler ultrasound will be obtained at month 3 as standard of care in all 
patients to evaluate residual vein thrombosis.  
 
10.0     Outpatient Visits 
 
Patients will be seen in the outpatient thrombosis clinic at least on enrollment, month 3, 
and month 9 (conclusion of the study), at each time side effects of drugs, symptoms of 
bleeding and recurrent VTE, post thrombotic syndrome will be assessed. Pill count and 
drug compliance will be assessed as well. Enrollees will be given $6 dollar food vouchers 
to be used in the cafeteria for each of the scheduled study visit (enrollment, 3 and 9 
months) to compensate their efforts of fasting prior to lab drawn.  
 
11.0    Safety Monitoring During Study 
 
11.1 
To ensure the safety of all patients enrolled on this study, careful monitoring and follow-
ups for all patients will be important aspects of this study. Patients will be monitored 
closely with laboratory and clinical evaluations at regular intervals defined in the study 
protocol. 
 
11.2     At each visit (enrollment, 3 and 9 months), patients will be evaluated by physicians and 
study personnel for side effects of statins and anticoagulation. The management of side 
effects from statins will follow the routine practice for patients who are prescribed with 
statins for other medical problems, and will be left to the discretion of the treating 
physicians, but the study team and PI will be available for any assistance if questions of 
the optimal management of adverse events were to arise. Live function tests will be 
obtained prior to and at each follow-up visit. Creatitine kinase (CK) test will not be 
collected without symptoms, but will be collected if patients were to develop symptoms 
of muscle pain or weakness. Patients with new onset of transaminitis or CK above three 
times of upper limit of normal or significant symptoms will be taken off statins. Their 
abnormal laboratory tests will be monitored at least every 2 weeks, more frequently if 
needed, until the abnormalities resolve. Patients will only resume statins if their 
symptoms resolve, blood tests normalize, and if patients are willing to resume treatment. 
Recurrent test abnormality or symptoms will require permanent discontinuation of 
statins. 
 
 
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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12.0  Adverse Events/Severe Adverse Events 
 
12.1    Definition: Adverse events as defined in the International Council on Harmonisation 
Guidelines for Good Clinical Practice are described as “an untoward medical occurrence 
in a patient or clinical investigation administered a pharmaceutical product that does not 
necessarily have a causal relationship with this treatment.”  
 
12.2     A serious adverse event is an adverse event that: 
 Is fatal 
 Is life threatening (places the subject at immediate risk of death) 
 Results in inpatient hospitalization or prolonged existing hospitalization 
 Results in persistent or significant disability/incapacity 
 Is a congenital anomaly/birth defect 
 Includes other significant hazard 
 
12.3      Grading of Adverse Events – Events will be graded by using the Common Terminology 
Criteria for Adverse Events (CTCAE) Criteria version 4.0. 
 
12.4       Attribution – the determination and documentation of whether an adverse event is related 
to a medical procedure. 
 
Attribution Categories: 
1. Not Related –Event clearly related to other factors (e.g., clinical state, other therapies; 
concomitant drugs) 
2. Possible Related – Sequence of event is compatible with study drug, device, or 
procedure, but could have been produced by other factors 
3. Probably Related - Sequence of event is compatible with study drug, or procedure and 
cannot be explained by other factors without much doubt 
4. Definitely Related - Sequence of event is compatible with study drug, or procedure 
and beyond doubt cannot be explained by other factors 
 
12.5  Reporting of Adverse Events: Adverse events must be recorded within 7 days of the 
occurrence using the “Adverse Event” form (see Appendix).  The principal investigator 
will review all adverse event forms every month or more frequently if needed. All serious 
or unexpected adverse events must be communicated to the principal investigator 
immediately and will be reported to the IRB within 24 hours of learning of the event’s 
occurrence by phone, email, or SAE for regardless of attribution. Using these 
mechanisms of rapid reporting, we will be able to rapidly evaluate and minimize the 
potential risks to patients from the occurrence of these events.  Subjects will be instructed 
to report any adverse event(s) during the study and post-study. The investigator will 
notify the IRB of any unanticipated death or adverse event occurring after a participant 
has discontinued or terminated study participation that may reasonably be related to the 
study.  
 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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12.6  Follow-up of Adverse Events: Patients experiencing an adverse event will continue to be 
followed and carefully monitored until the resolution of the adverse event.  This applies 
whether the patient continues on study or is removed from the study for any reason. 
 
12.7  While no formal stopping rules are deemed necessary due to the well-known safety and 
tolerability of statins, the PI will be informed of any SAE immediately, determine 
correlation, and report to IRB.  The PI is responsible to continuously evaluate the safety 
for study continuation based on regular review of AE reports.  
 
13.0    Sample Procurement  
 
 
Both plasma and blood samples will be taken as a part of this protocol.  Theses samples 
will be obtained at the same time that standard clinical samples or procedures are being 
performed so patients will not be exposed to additional venipunctures or procedures by 
their participation in this study.   
 
14.0     Sample Storage and Handling 
  
 
At the time of enrollment and registration with the coordinating institution, arrangements 
will be made for the research blood samples to be delivered to the laboratory of Dr. Bryce 
Kerlin, Dr. Mark Hall, and Dr. Kenneth Riedl for processing and analysis. Research 
blood samples will be labeled with a unique identification number so that the data 
obtained may be correlated with the patient’s clinical course by the investigators at the 
end of the study, while the laboratory personnel performing the research testing will not 
be able to identity each individual patient.  This will help ensure patient confidentiality, 
as the investigators will be the only people to know the patient name and medical record 
number that correspond with the identification number.  Samples to be used for future 
study will be frozen at -80C and be stored in the benign hematology research office at 
the James Cancer Hospital, and in the laboratory of Dr. Kerlin, depending on the 
designated research testing to be done: thrombin generation assays will be performed in 
Dr. Kerlin’s laboratory at Nationwide Children’s. Inflammatory markers will be done in 
Dr. Mark Hall’s laboratory at Nationwide Children’s and lipidomic profiles will be 
performed at Dr. Kenneth Riedl’s laboratory at Ohio State. At the end of the study, at 
least one aliquot of each plasma and RNA samples will be delivered to Dr. Alan Mast’s 
laboratory at the Blood Center of Wisconsin for testing including TF, TFPI, and other 
clotting factors. Samples will be sent on dry ice via commercial carriers overnight.  
 
15.0    Risks Associated with Testing of Patient Samples 
 
 
A breach of confidentiality is the most serious risk which patients may be exposed to 
regarding to research testing of biologic samples. Confidentiality will be maintained by 
labeling the patient samples with only an identification number and not their name or 
medical record number.  By using this approach, patient samples may be tied to clinical 
information by the investigators but will preserve the confidentiality of patients.  Only the 
principal investigator, co-investigator, and the study nurse/coordinator will have access to 
both the study number and the corresponding patient identification information.  Clinical 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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information or laboratory data used for publication will not identify patients in any way 
that could potentially breach patient confidentiality.  The samples sent to BloodCenter of 
Wisconsin will be coded and all PHI will be removed so the laboratory personnel there 
will have no access to the PHI. The samples need to be coded and not de-identified, so 
the principal and co-investigators can correlate clinical with research laboratory findings. 
Even though more research laboratory tests are planned in this protocol amendment, it 
will be accomplished within the same amount of blood that was originally planned to be 
drawn and therefore expose no additional risks to subsequent enrollees.  
 
16.0   Thrombin generation assay 
 
Blood samples for measurement of thrombin generation will be drawn in vacutainer tubes 
containing 3.2% sodium citrate with and without corn trypsin inhibitor (CTI) and be 
processed appropriately within an hour prior to storage. Blood samples will be 
centrifuged at 2500g (relative centrifugal force) for 15 minutes (min). Plasma will be 
transferred into plastic tubes, and re-centrifuged at 2500g for 15 min. Plasma will be 
decanted again after second spin and stored in aliquots of 0.2 mL (platelet poor plasma). 
The platelet poor plasma will then be snap frozen and stored at -70°C until testing as bulk 
at the end of the study. 
The thrombin generation assays will be performed using a commercial kit 
(TECHNOTHROMBIN® TGA, Technoclone, Vienna, Austria) according to the 
manufacturer’s instructions in Dr. Kerlin’s laboratory. The assays are performed based on 
monitoring the fluorescence generated by thrombin cleavage of a fluorogenic substrate 
over time, after the activation of the coagulation cascade by tissue factor. At least two 
reagents will be used. Parameters that will be recorded include: lag phase, time to peak 
thrombin concentration, peak thrombin concentration, and endogenous thrombin potential 
(measured by area under the curve). 
 
17.0    Inflammatory marker assessment 
 
Serum IL-6, IL-8, and TNF-α levels will be measured using commercial enzyme-linked 
immunosorbent assay (ELISA) according to the manufacturer’s instructions in Dr. 
Mark Hall’slaboratory. 
 
18.0    Lipodomic profiles 
 
18.1    Lipidomic profiles will be done in Dr. Riedl’s laboratory. Blood samples for measurement 
of lipidomic profiles will be drawn in two vacutainer tubes, one containing 3.2% sodium 
citrate and the other being a serum separator tube.  
 
After collection, blood will be separated into individual constituents including 
serum. Lipidomic profiles will be collected on blood samples using a triple quadrupole 
mass spectrometry platform called Lipidyzer which is enabled through ion mobility 
hardware. This assay covers 13 different lipid classes and uses 56 stable isotope internal 
standards to provide quantitative results for as many as 1,100 lipid species. A separate 
LCMS panel will be utilized to determine eicosanoid profiles in blood. Both of these 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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assays will be conducted in the Nutrient & Phytochemical Analytic Shared Resource 
(NPASR) of the OSU Comprehensive Cancer Center under the direction of Acting 
Director, Ken Riedl..  
 
19.0    Plasma levels and RNA expression of tissue factor (TF), tissue factor pathway 
inhibitor (TFPI), and other clotting factors 
 
19.1    These tests will be done in Dr. Alan Mast’s laboratory at BloodCenter of Wisconsin 
according to their standard protocol. The plasma samples will be frozen and stored at -
80°C at the Ohio State University until the end of the study, when samples will be 
delivered to Dr. Mast’s lab to perform the research testing. 
 
19.2     After plasma was removed from each tube as per protocol above, the buffy coats in the 
tubes will be collected as much as possible and stored in RNAlater® reagent as per 
manufacturer’s protocol. These buffy coats will be used for analysis of RNA expression in 
Dr. Mast’s laboratory at the end of the study, according to their standard protocol.  
 
20.0  Statistical Analysis 
 
 
At least 66 evaluable patients will be randomized (1:1 allocation, 33 evaluable patients 
in each arm) into “anticoagulation” versus “anticoagulation +atorvastatin” arms. The 
primary endpoints are thrombin peak and thrombin generation. Based on the published 
data,8,9 we anticipate an 80% power to detect an effect size of 0.7 standard deviations in 
the reduction of thrombin peak or thrombin generation between the two arms with two-
side alpha of 0.05 using a two-sample t test. An overall alpha of 0.1 for this pilot study 
was split into 0.05 for each endpoint (thrombin peak and thrombin generation) using 
Bonferroni method. Considering possible drop-outs, our target enrollment will be 80 
patients, with 40 on each arm. 
 
All data will be collected on study-approved data collection forms and entered into 
Research Electronic Data Capture (REDCap), which is a secure, web-based application 
designed to support data capture for research studies by building and managing online 
surveys and databases. The system was built with the help of the OSU Research 
Informatics Services. 
 
Descriptive statistics (i.e. means, standard deviations, 95% confidence intervals for 
continuous variables, and frequencies for discrete data) and graphical analyses will be 
used for all clinical outcomes, demographics, VTE related events, laboratory results, and 
adverse events for each arm. The secondary endpoints will be compared between the 
two arms using chi- square test or Fisher’s exact test, whichever is appropriate. The 
associations between treatment type and the change in biomarkers will be evaluated 
using two-sample t test. Log-transform will be used if necessary. Linear mixed models 
will be used to study the associations between the primary endpoints and treatment, and 
trend across time. The interaction term “arm x time” will be included in the model to 
study whether the two treatment arms have different “behavior” across time. Potential 
confounders will be included in the models. Thrombin peak concentration and 

A Pilot Study of Using Statins in Patients with Acute Venous Thromboembolism (VTE) 
IRB Protocol Number: 2014H0262 
 
Approval Date: 11/15/2017 
 
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endogenous thrombin potential at 9 months will be compared between the patients on 
anticoagulation and those off anticoagulation. 
 
21.0     Costs of the Study 
 
 
Given the fact that all patients will be receiving standard therapy for VTE 
(anticoagulation), either the patient or their insurance carrier will be responsible for all of 
the medical expenses associated with the treatment and follow up of their VTE. The costs 
associated with the research laboratory studies will be paid by the study researchers and 
will not be passed on to patients or their insurance carriers. The cost of atorvastatin will 
be reimbursed by the study researchers. Patients enrolled on this study will not incur any 
additional expenses as a result of enrolling on this protocol. 
 
22.0 
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IRB Protocol Number: 2014H0262 
 
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