Pulmonary Disease, Chronic ObstructiveNCT01745419
Prevalence of Different Haptoglobin Phenotypes in Patients With COPD- Frequent Exacerbators Versus Non Exacerbators
An observational study of Pulmonary Disease, Chronic Obstructive, run by Carmel Medical Center at 1 site in Israel.
Trial summary
Chronic obstructive pulmonary disease (COPD) is a common disease in smokers. COPD has a slowly deteriorating course, punctuated by exacerbations- acute events characterized by increasing shortness of breath and putrid sputum. Exacerbations of COPD may be precipitated by several factors, most commonly infections. Read more about this study
- Who is it for?
- Ages 40 to 90 years
- Conditions the study is for
- Pulmonary Disease, Chronic Obstructive
- Type of treatment
- Pill or oral medicine
- How many people
- 105 participants
- Where
- 1 siteIsrael
- Study dates
- Jan 2013to Jun 2016
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 40 to 90 years
- All sexes
- No healthy volunteers
- Adult, Older adult
Study population: Patients with COPD
Who may be able to join
- Patients diagnosed with COPD according to GOLD guidelines1
- Age 40-100 years
- Smoking history of > 10 pack - years
- Spirometry consistent with airflow obstruction (FEV1/FVC ratio < 70%)
- Moderate to severe airflow obstruction (FEV1<60%)
- At least two episodes of acute exacerbations in the former 12 months, or no exacerbations in the former 24 months. (Acute exacerbations are defined as worsening symptoms requiring treatment with systemic steroids (oral or parenteral) or antibiotics, a visit to the emergency room, and/or admission to a hospital. Events separated by at least 21 days are considered as separate events of exacerbation.)
Who cannot join
- Patients with a history of any of the following conditions will be excluded from the study:
- Active Tuberculosis
- Pulmonary fibrosis or Asbestosis
- Organ transplantation
- Lung volume reduction surgery
- Previous lung or lobe resection.
Trial locations and contacts
1 site in 1 country.
About this study
Chronic obstructive pulmonary disease (COPD) is a common disease in smokers. COPD has a slowly deteriorating course, punctuated by exacerbations- acute events characterized by increasing shortness of breath and putrid sputum. Exacerbations of COPD may be precipitated by several factors, most commonly infections.
Exacerbation frequency generally increases with declining lung function. However, some patients with COPD consistently experience a higher rate of exacerbations than others despite similar severity of COPD. This has led researchers to postulate the existence of a distinct subgroup of "frequent exacerbators" . Recent work has also brought attention to a subset of patients who experience remarkably few exacerbations despite significantly impaired lung function. Careful characterization of both of these extreme subgroups of COPD may offer additional insights into why certain patients are prone to frequent exacerbations while others remain relatively protected.
Read more
Haptoglobin (Hp) is a protein produced predominately by the liver . In humans two types of genes for Hp exist (1 and 2) with possible combinations of these two genes- 1-1, 1-2, or 2-2. The Hp 2 gene is believed to have arisen from the Hp 1 gene in human evolution. Subsequently the prevalence of the Hp 2 allele has spread throughout the world, probably as a result of its ability to provide a selective advantage against infectious disease. The Hp 1-2 combination is a very common one. In most western countries, the prevalence of the Hp genotypes is 16% Hp 1-1, 36% Hp 2-2 and 48% Hp 2-1.
The Hp gene form has been shown to be associated with disease. Specifically, Hp phenotypes have been found to affect propensity to atherosclerosis in Diabetic individuals. There have been several studies suggesting that the Hp 2-2 phenotype is associated with a protection against infectious complications.
In view of the importance of respiratory infections on COPD exacerbations, and of the gained knowledge of Haptoglobin subtypes on propensity to infection, we propose to investigate whether Haptoglobin subtypes are in correlation with the "frequent exacerbator" phenotype of COPD. We postulate that, since people with Hp 1-1 are more prone to infection, the frequency of the Hp 1-1 phenotype will be higher in "frequent exacerbators" of COPD than in "non- exacerbators".
To test our hypothesis we propose to determine Hp phenotype in two groups of COPD patients: one with frequent exacerbations and one with no exacerbations, and compare the relative frequency of the 1-1 phenotype in the two groups.
Read the detailed description
Laboratory Procedures:
Overview:
3 ml of Serum will be collected and stored at 4 degrees Celsius (up to one week) or minus 20 degrees Celsius until delivery to the laboratory.
Haptoglobin phenotype will be determined at the Department of Anatomy and Cell Biology, the B. Rappaport Faculty of Medicine, 1 Efron st. Haifa, Israel. The method for phenotypin Hp is protein gel electrophoresis.
Serum remaining will be kept frozen in minus 70 degrees Celsius for 5 years in a freezer shelf dedicated for the Pulmonology Institute at the Serology Laboratory in Carmel Medical Center for future analysis of proteins.
Haptoglobin Electrophoresis:
The Haptoglobin protein is separated based on size. Hp 2-2 is the largest therefore was expected to travel the shortest distance in the gel and Hp 1-1 is the smallest and therefore would travel the farthest down the gel. The gel that is used for the electrophoresis was a Polyacrylamide gel. Two concentrations are used in order to provide loading and separating conditions.
The lower gel is a Polyacrilamide gel that contains 10.8 ml 1M Tris pH 8.8, 3.55 ml of Acrylamide 40%, 11.5 ml of dH2O, 225 μl of Ammonium persulfate (APS) 10%, and 18 μl of tetramethylethylenediamine (TEMED). The upper gel is a Polyacrylamide gel that contains 937.5 μl of 1M Tris pH 6.8, 787.5 μl of Acrylamide 40%, 5.7 ml of dH2O, 75 μl of APS 10%, and 7.5 μl of TEMED.
The lower gel is poured in between two glass plates and left for 30 minutes to solidify. Once lower gel is solidified upper gel is poured on top of it, and left for 30 min to solidify. Wells comb placed superiorly to upper gel in order to create wells for sample placement.
The sample for each patient is prepared using 10 μl of the serum, 2 μl of Hemoglobin, and 12 μl of a commercial loading buffer. The sample is then carefully placed in the prepared wells using a pipette. 4 wells are reserved for 3 controls of Hp 2-2, Hp 2-1, and Hp 1-1 followed medially by an empty well.
The western blot analysis is run in a Protean II xi cell by BIO-RAD. The apparatus is assembled and an electrophoresis running buffer is added. The running buffer contains 15.1 g of Tris, 72 g of Glycine, and 1000 ml of DDW. The running buffer was then placed in the upper and lower reservoir of the Protean II. The electrophoresis was run at 240 Volts for 3 hrs.
Once the samples are finished running, the gel is removed from between the glass plates. The gel was then stained using a color staining solution. The Solution was made using 40 mg of 3,3'-5,5' tetramethyl benzidine and 20 ml of methanol. This was left to mix for 15 minutes. Next 2 ml of dimethyl sulfoxide is added and left to mix for 5 min. This was followed by the addition of 40 ml of acetic acid 5% which is made of 2 ml AA and 38 ml of DDW. Next 40 mg of potassium fericyanide dissolved in 4 ml of DDW are added. Finally 600 μl of H2O2 is added to the staining solution and left to mix for 10 min.
Once the staining solution is ready, the gel is placed in a 22 cm by 32 cm tray and the staining solution is poured over the gel. The tray is then placed on a shaker for 25 minutes. Once the Gel is finished being stained it is legible. Nevertheless, the gel is photographed for image enhancement and data storage. The gel is removed from the tray and placed in an Image Analyzer (Imagequant LAS 4000) for imaging, or Canon powershot sx40 hs digital camera.
Data Tabulation:
The data - the Haptoglobin phenotype with corresponding IDs - is gathered and tabulated into a Microsoft excel table.
- Conditions
- Pulmonary Disease, Chronic Obstructive
- Keywords
- COPD, Exacerbation, Haptoglobin
- Medical subject headings
- Pulmonary Disease, Chronic Obstructive
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
| Study group | Role | What they receive |
|---|---|---|
| COPD Frequent Exacerbators | COPD patients having experienced at least two episodes of acute exacerbations in the former 12 months. (Acute exacerbations are defined as worsening symptoms requiring treatment with systemic steroids (oral or parenteral) or antibiotics, a visit to the emergency room, and/or admission to a hospital. Events separated by at least 21 days are considered as separate events of exacerbation.) | |
| COPD non- exacerbators | COPD patients who have not experienced exacerbations in the former 24 months. (Acute exacerbations are defined as worsening symptoms requiring treatment with systemic steroids (oral or parenteral) or antibiotics, a visit to the emergency room, and/or admission to a hospital. Events separated by at least 21 days are considered as separate events of exacerbation.) |
What is being measured
The primary outcome is the main question the study is designed to answer.
Prevalence of Hp 1-1 phenotype
Prevalence as percentage of Hp 1-1 phenotypes in each group of patients
Measured: 1 visit- approximately 2 hours.
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted10 Dec 2012 (estimated)
- Study startJan 2013
- Primary completionJan 2015
- Study completionJun 2016
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Pulmonary Disease, Chronic ObstructiveStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case-control
- Time perspective
- Retrospective
- Enrolment
- 105 participants (actual)
Sponsor and investigators
- Lead sponsor
- Carmel Medical Center
- Sponsor type
- University, hospital or other
- Collaborators
- Technion, Israel Institute of Technology
- Responsible party
- Principal investigator, Michal Steinberg, Carmel Medical Center
- Investigators
- Michal Shteinberg, MD, PhD, Principal investigator, Pulmonology Institute, Carmel Medical Center
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT01745419
- Sponsor's protocol ID
- CMC-12-0121-CTIL
- Status last verified
- Jul 2016
- First submitted
- 5 Dec 2012
- Last updated
- 28 Jul 2016
- Results posted
- No
- Data monitoring committee
- No
Study at a glance
From the registry record, last updated by the sponsor on 28 Jul 2016. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Prevalence of Different Haptoglobin Phenotypes in Patients With COPD- Frequent Exacerbators Versus Non Exacerbators
- ClinicalTrials.gov ID
- NCT01745419
- Acronym / sponsor ID
- CMC-12-0121-CTIL
- Conditions
- Pulmonary Disease, Chronic Obstructive
- Keywords
- COPD, Exacerbation, Haptoglobin
- Study type
- Observational
- Recruitment status
- Completed
- Enrolment
- 105 participants (actual)
- Lead sponsor
- Carmel Medical CenterUniversity, hospital or other
- Collaborators
- Technion, Israel Institute of Technology
- Study start
- Jan 2013
- Primary completion
- Jan 2015 (actual)
- Study completion
- Jun 2016 (actual)
- First posted
- 10 Dec 2012 (estimated)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
Chronic obstructive pulmonary disease (COPD) is a common disease in smokers. COPD has a slowly deteriorating course, punctuated by exacerbations- acute events characterized by increasing shortness of breath and putrid sputum. Exacerbations of COPD may be precipitated by several factors, most commonly infections.
Exacerbation frequency generally increases with declining lung function. However, some patients with COPD consistently experience a higher rate of exacerbations than others despite similar severity of COPD. This has led researchers to postulate the existence of a distinct subgroup of "frequent exacerbators" . Recent work has also brought attention to a subset of patients who experience remarkably few exacerbations despite significantly impaired lung function. Careful characterization of both of these extreme subgroups of COPD may offer additional insights into why certain patients are prone to frequent exacerbations while others remain relatively protected.
Haptoglobin (Hp) is a protein produced predominately by the liver . In humans two types of genes for Hp exist (1 and 2) with possible combinations of these two genes- 1-1, 1-2, or 2-2. The Hp 2 gene is believed to have arisen from the Hp 1 gene in human evolution. Subsequently the prevalence of the Hp 2 allele has spread throughout the world, probably as a result of its ability to provide a selective advantage against infectious disease. The Hp 1-2 combination is a very common one. In most western countries, the prevalence of the Hp genotypes is 16% Hp 1-1, 36% Hp 2-2 and 48% Hp 2-1.
The Hp gene form has been shown to be associated with disease. Specifically, Hp phenotypes have been found to affect propensity to atherosclerosis in Diabetic individuals. There have been several studies suggesting that the Hp 2-2 phenotype is associated with a protection against infectious complications.
In view of the importance of respiratory infections on COPD exacerbations, and of the gained knowledge of Haptoglobin subtypes on propensity to infection, we propose to investigate whether Haptoglobin subtypes are in correlation with the "frequent exacerbator" phenotype of COPD. We postulate that, since people with Hp 1-1 are more prone to infection, the frequency of the Hp 1-1 phenotype will be higher in "frequent exacerbators" of COPD than in "non- exacerbators".
To test our hypothesis we propose to determine Hp phenotype in two groups of COPD patients: one with frequent exacerbations and one with no exacerbations, and compare the relative frequency of the 1-1 phenotype in the two groups.
Detailed description
Laboratory Procedures:
Overview:
3 ml of Serum will be collected and stored at 4 degrees Celsius (up to one week) or minus 20 degrees Celsius until delivery to the laboratory.
Haptoglobin phenotype will be determined at the Department of Anatomy and Cell Biology, the B. Rappaport Faculty of Medicine, 1 Efron st. Haifa, Israel. The method for phenotypin Hp is protein gel electrophoresis.
Serum remaining will be kept frozen in minus 70 degrees Celsius for 5 years in a freezer shelf dedicated for the Pulmonology Institute at the Serology Laboratory in Carmel Medical Center for future analysis of proteins.
Haptoglobin Electrophoresis:
The Haptoglobin protein is separated based on size. Hp 2-2 is the largest therefore was expected to travel the shortest distance in the gel and Hp 1-1 is the smallest and therefore would travel the farthest down the gel. The gel that is used for the electrophoresis was a Polyacrylamide gel. Two concentrations are used in order to provide loading and separating conditions.
The lower gel is a Polyacrilamide gel that contains 10.8 ml 1M Tris pH 8.8, 3.55 ml of Acrylamide 40%, 11.5 ml of dH2O, 225 μl of Ammonium persulfate (APS) 10%, and 18 μl of tetramethylethylenediamine (TEMED). The upper gel is a Polyacrylamide gel that contains 937.5 μl of 1M Tris pH 6.8, 787.5 μl of Acrylamide 40%, 5.7 ml of dH2O, 75 μl of APS 10%, and 7.5 μl of TEMED.
The lower gel is poured in between two glass plates and left for 30 minutes to solidify. Once lower gel is solidified upper gel is poured on top of it, and left for 30 min to solidify. Wells comb placed superiorly to upper gel in order to create wells for sample placement.
The sample for each patient is prepared using 10 μl of the serum, 2 μl of Hemoglobin, and 12 μl of a commercial loading buffer. The sample is then carefully placed in the prepared wells using a pipette. 4 wells are reserved for 3 controls of Hp 2-2, Hp 2-1, and Hp 1-1 followed medially by an empty well.
The western blot analysis is run in a Protean II xi cell by BIO-RAD. The apparatus is assembled and an electrophoresis running buffer is added. The running buffer contains 15.1 g of Tris, 72 g of Glycine, and 1000 ml of DDW. The running buffer was then placed in the upper and lower reservoir of the Protean II. The electrophoresis was run at 240 Volts for 3 hrs.
Once the samples are finished running, the gel is removed from between the glass plates. The gel was then stained using a color staining solution. The Solution was made using 40 mg of 3,3'-5,5' tetramethyl benzidine and 20 ml of methanol. This was left to mix for 15 minutes. Next 2 ml of dimethyl sulfoxide is added and left to mix for 5 min. This was followed by the addition of 40 ml of acetic acid 5% which is made of 2 ml AA and 38 ml of DDW. Next 40 mg of potassium fericyanide dissolved in 4 ml of DDW are added. Finally 600 μl of H2O2 is added to the staining solution and left to mix for 10 min.
Once the staining solution is ready, the gel is placed in a 22 cm by 32 cm tray and the staining solution is poured over the gel. The tray is then placed on a shaker for 25 minutes. Once the Gel is finished being stained it is legible. Nevertheless, the gel is photographed for image enhancement and data storage. The gel is removed from the tray and placed in an Image Analyzer (Imagequant LAS 4000) for imaging, or Canon powershot sx40 hs digital camera.
Data Tabulation:
The data - the Haptoglobin phenotype with corresponding IDs - is gathered and tabulated into a Microsoft excel table.
Study design
- Observational model
- Case-control
- Time perspective
- Retrospective
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| COPD Frequent Exacerbators | – | –COPD patients having experienced at least two episodes of acute exacerbations in the former 12 months. (Acute exacerbations are defined as worsening symptoms requiring treatment with systemic steroids (oral or parenteral) or antibiotics, a visit to the emergency room, and/or admission to a hospital. Events separated by at least 21 days are considered as separate events of exacerbation.) |
| COPD non- exacerbators | – | –COPD patients who have not experienced exacerbations in the former 24 months. (Acute exacerbations are defined as worsening symptoms requiring treatment with systemic steroids (oral or parenteral) or antibiotics, a visit to the emergency room, and/or admission to a hospital. Events separated by at least 21 days are considered as separate events of exacerbation.) |
Who may qualify
- Ages
- 40 to 90 years
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Adult, Older adult
- Study population
- Patients with COPD
Key inclusion criteria (6 listed)
- Patients diagnosed with COPD according to GOLD guidelines1
- Age 40-100 years
- Smoking history of > 10 pack - years
- Spirometry consistent with airflow obstruction (FEV1/FVC ratio < 70%)
- Moderate to severe airflow obstruction (FEV1<60%)
Key exclusion criteria (6 listed)
- Patients with a history of any of the following conditions will be excluded from the study:
- Active Tuberculosis
- Pulmonary fibrosis or Asbestosis
- Organ transplantation
- Lung volume reduction surgery
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Primary outcome
Prevalence of Hp 1-1 phenotype
Time frame: 1 visit- approximately 2 hours.
Prevalence as percentage of Hp 1-1 phenotypes in each group of patients
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 28 Jul 2016.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Michal Shteinberg, MD, PhD principal investigator
Pulmonology Institute, Carmel Medical Center
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Pulmonary Disease, Chronic Obstructive, run by Carmel Medical Center at 1 site in Israel.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 6 inclusion and 6 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 28 Jul 2016. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Israel. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Prevalence of Hp 1-1 phenotype (time frame: 1 visit- approximately 2 hours.). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 28 Jul 2016.