Evaluating the Vitamin K2 Status of Calcium-based Stone Formers

Study Overview
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Brief Summary
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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.

Detailed Description
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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.

Official Title
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Evaluating the Vitamin K2 Status of Calcium-based Stone Formers

Conditions
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Kidney Calculi, Calcium Oxalate Kidney Stones, Vitamin K 2, Human, Kidney Stone, Calcium Oxalate Urolithiasis, Calcium Phosphate Urolithiasis, Nephrolithiasis

Participation Criteria
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Eligibility Criteria
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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.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Stone Formers<br>Individuals who have experienced at least one incidence of calcium-based kidney stones in the last 12 months | |
| Controls<br>Individuals who have never had a kidney stone in their lifetime. | |

What is the study measuring?
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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 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Kidney stone disease, Microbiota, Menaquinone, Vitamin K2, Matrix Gla protein, dp-ucMGP, Nephrolithiasis