Evaluating the Vitamin K2 Status of Calcium-based Stone Formers

This is an observation, single site-study with one study visit during which all data and samples will be collected. Study participants will be asked to provide blood, urine, and fecal samples so that the investigators may study the differences in the gut microbiota, vitamin K2 levels, and other parameters between participants who form kidney stones and those who do not.

Study Overview

• Status: Active, not recruiting
• Conditions: Kidney Calculi; Nephrolithiasis; Kidney Stone; Human; Calcium Oxalate Urolithiasis; Vitamin K 2; Calcium Oxalate Kidney Stones; Calcium Phosphate Urolithiasis

Detailed Description

It is hypothesized that calcium-based stone formers will have an altered fecal gut microbiota compared to non-stone former controls. This altered microbiota will have a lower abundance of bacteria that produce menaquinones (vitamin K2), thus stone formers will also have a different blood menaquinone profile compared to controls. Ultimately, the different levels of menaquinones will result in increased inactive Matrix Gla protein (dp-ucMGP), which is a key protein that sequesters free calcium. To test this hypothesis, calcium-based stone former and non-stone forming controls will be recruited to a single site, observation study to collect urine, blood, and fecal samples. These samples will be used to determine dp-ucMGP levels, menaquinone profiles, the composition of the gut microbiota, and other parameters of interest.

• Study Type: Observational
• Enrollment (Actual): 60

Contacts and Locations

• Study Contact: Name: Jennifer Bjazevic, MD; Phone Number: 66036 519-646-6100; Email: Jennifer.Bjazevic@sjhc.london.on.ca
• Study Contact Backup: Name: John A Chmiel, MSc; Phone Number: 905-912-2382; Email: jchmiel4@uwo.ca

Study Locations

• Canada
  • Ontario
    • London, Ontario, Canada, N6J 3T9
      • St. Joseph's Health Care London

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 65 years (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Stone formers will be recruited from the Urology clinic or London and surrounding community. Controls will be recruited from the London and surrounding community.

Description

Inclusion Criteria:

• Male/Female, 18 - 65 years old
• No self-reported kidney stones during their lifetime (controls)
• Ultrasound examination confirming absence of kidney stones (controls)
• Have had at least 1 incidence of a clinically confirmed calcium-based kidney stone in the last 12 months (stone formers)
• Ability to collect a clean catch urine sample
• Prescription and over-the-counter drugs unchanged for ≥30 days
• Willingness to provide medical information, blood, urine, and fecal samples

Exclusion Criteria:

• Current, or within 30 days, use of antibiotics or antifungals
• Current, or within 30 days, use of vitamin K antagonists
• Current probiotic use or any use within 14 days of screening sample collection should be recorded
• A history or currently undergoing immunosuppressive drug therapy, chemotherapy, or radiation therapy 2021-06-29 1.0 Page 4 of 6
• Fecal incontinence
• History of disorder with abnormal calcium regulation such as hyperparathyroidism, active malignancy, or osteoporosis.

Study Plan

How is the study designed?

Design Details

• Observational Models: Case-Control
• Time Perspectives: Cross-Sectional

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort
Stone Formers; Individuals who have experienced at least one incidence of calcium-based kidney stones in the last 12 months
Controls; Individuals who have never had a kidney stone in their lifetime.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Fecal microbiota composition of stone-formers and controls	Fecal samples will be collected using out validated toilet paper method. Microbial DNA will be extracted and sequenced using next-generation sequencing.	At baseline only
Concentration of urine dp-ucMGP (dephosphorylated-uncarboxylated Matrix Gla Protein)	dp-ucMGP will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of blood dp-ucMGP (dephosphorylated-uncarboxylated Matrix Gla Protein)	dp-ucMGP will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of blood total osteocalcin (OC)	Total OC will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of blood undercarboxylated osteocalcin (ucOC)	ucOC will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of urine total osteocalcin (OC)	Total OC will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of urine undercarboxylated osteocalcin (ucOC)	ucOC will be quantified using an enzyme-linked immunosorbent assay	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-4	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-7	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-8	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-9	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-10	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-11	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-12	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only
Concentration of blood menaquinones (vitamin K2) - MK-13	Menaquinones will be quantified using liquid chromatography with mass spectrometry or fluorescence.	At baseline only

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Concentration of blood fetuin A	Fetuin A will be quantified using enzyme-linked immunosorbent assay	At baseline only
Concentration of urine fetuin A	Fetuin A will be quantified using enzyme-linked immunosorbent assay	At baseline only
Percentage of blood Hemoglobin A1C (HbA1c)	HbA1c will be quantified in the core laboratory as per established protocols	At baseline only
Total plasma calcium	Calcium levels will be quantified in the core laboratory	At baseline only
Concentration of ionized calcium in blood	Calcium levels will be quantified in the core laboratory	At baseline only
Concentration of blood albumin	Albumin levels will be quantified in the core laboratory as per established protocols	At baseline only
Concentration of urinary γ-carboxyglutamic acid	γ-carboxyglutamic acid will be quantified using high-performance liquid chromatography and normalized to creatinine	At baseline only
Concentration of urinary creatinine	Creatinine will be quantified using high-performance liquid chromatography	At baseline only
Concentration of urinary oxalate	Oxalate will be quantified using high-performance liquid chromatography and normalized to creatinine	At baseline only
Concentration of urinary phosphate	Phosphate will be quantified in the core laboratory as per established protocols	At baseline only

Collaborators and Investigators

• Sponsor: Lawson Health Research Institute
• Collaborators: St. Joseph's Health Care London
• Investigators: Principal Investigator: Jennifer Bjazevic, MD, Lawson Heath Research

Publications and helpful links

General Publications

• Scales CD Jr, Smith AC, Hanley JM, Saigal CS; Urologic Diseases in America Project. Prevalence of kidney stones in the United States. Eur Urol. 2012 Jul;62(1):160-5. doi: 10.1016/j.eururo.2012.03.052. Epub 2012 Mar 31.
• Fraser JD, Price PA. Lung, heart, and kidney express high levels of mRNA for the vitamin K-dependent matrix Gla protein. Implications for the possible functions of matrix Gla protein and for the tissue distribution of the gamma-carboxylase. J Biol Chem. 1988 Aug 15;263(23):11033-6.
• Moe OW. Kidney stones: pathophysiology and medical management. Lancet. 2006 Jan 28;367(9507):333-44. doi: 10.1016/S0140-6736(06)68071-9.
• Stanford J, Charlton K, Stefoska-Needham A, Ibrahim R, Lambert K. The gut microbiota profile of adults with kidney disease and kidney stones: a systematic review of the literature. BMC Nephrol. 2020 Jun 5;21(1):215. doi: 10.1186/s12882-020-01805-w.
• Conly J, Stein K. Reduction of vitamin K2 concentrations in human liver associated with the use of broad spectrum antimicrobials. Clin Invest Med. 1994 Dec;17(6):531-9.
• Sato T, Schurgers LJ, Uenishi K. Comparison of menaquinone-4 and menaquinone-7 bioavailability in healthy women. Nutr J. 2012 Nov 12;11:93. doi: 10.1186/1475-2891-11-93.
• Schurgers LJ, Vermeer C. Determination of phylloquinone and menaquinones in food. Effect of food matrix on circulating vitamin K concentrations. Haemostasis. 2000 Nov-Dec;30(6):298-307. doi: 10.1159/000054147.
• Schurgers LJ, Vermeer C. Differential lipoprotein transport pathways of K-vitamins in healthy subjects. Biochim Biophys Acta. 2002 Feb 15;1570(1):27-32. doi: 10.1016/s0304-4165(02)00147-2.
• Wei FF, Thijs L, Zhang ZY, Jacobs L, Yang WY, Salvi E, Citterio L, Cauwenberghs N, Kuznetsova T, E A Drummen N, Hara A, Manunta P, Li Y, Verhamme P, Allegaert K, Cusi D, Vermeer C, Staessen JA. The risk of nephrolithiasis is causally related to inactive matrix Gla protein, a marker of vitamin K status: a Mendelian randomization study in a Flemish population. Nephrol Dial Transplant. 2018 Mar 1;33(3):514-522. doi: 10.1093/ndt/gfx014.
• Chmiel JA, Stuivenberg GA, Al KF, Akouris PP, Razvi H, Burton JP, Bjazevic J. Vitamins as regulators of calcium-containing kidney stones - new perspectives on the role of the gut microbiome. Nat Rev Urol. 2023 May 9:1-23. doi: 10.1038/s41585-023-00768-5. Online ahead of print.

Study record dates

Study Major Dates

• Study Start (Actual): August 1, 2022
• Primary Completion (Actual): June 8, 2023
• Study Completion (Estimated): October 1, 2023

Study Registration Dates

• First Submitted: September 22, 2021
• First Submitted That Met QC Criteria: October 5, 2021
• First Posted (Actual): October 18, 2021

Study Record Updates

• Last Update Posted (Actual): June 28, 2023
• Last Update Submitted That Met QC Criteria: June 27, 2023
• Last Verified: June 1, 2023

More Information

Terms related to this study

• Keywords: Microbiota; Nephrolithiasis; Vitamin K2; Matrix Gla protein; Kidney stone disease; Menaquinone; dp-ucMGP
• Additional Relevant MeSH Terms: Kidney Diseases; Urologic Diseases; Pathological Conditions, Anatomical; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Male Urogenital Diseases; Urinary Calculi; Urolithiasis; Calculi; Kidney Calculi; Nephrolithiasis
• Other Study ID Numbers: 119537

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No