Ignite Creation Date:
2024-05-05 @ 4:53 PM
Last Modification Date:
2024-10-26 @ 9:25 AM
Study NCT ID:
NCT00331942
Status:
UNKNOWN
Last Update Posted:
2007-03-02
First Post:
2006-05-27
Brief Title:
Breast Cancer Associated Antibodies
Sponsor:
Lab Discoveries Ltd
Organization:
Lab Discoveries Ltd
Study Overview
Official Title:
Study of Breast Cancer Associated Antibodies
Status:
UNKNOWN
Status Verified Date:
2006-04
Last Known Status:
RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
To determine the ability of the Cimmunology process to lead to in vitro antibody production the ability of the ELISA assays to detect any of those antibodies and to establish the relationship between the ELISA results and the clinical pathological status of the patient The statistical significance of the CAAb test results will be determined
Detailed Description:
General Information
Antibodies are a specific response to any foreign antigen and they are usually detectable in the serum within 5-7 days after the initial exposure to it However there are some cases where there is a suppression of the specific immune response such as in the case of a tumor To date there are no reports on serum antibodies that are associated to cancer Some tumors when surgically removed and studied have been found to have both infiltrates of T lymphocytes and antibodies bound to the tumor cells These antibodies and other specific immune responses are assumed to be formed at later stages of the tumor growth however even in those cases serum antibodies have not been reported probably due to them being mostly absorbed by the tumor mass
Since the mutations of a normal cell that lead to a malignant cell mass involves changes in the structure and function of the cells it could be assumed that the immune system has recognized some of these changes as antigenic determinates that should be responded against if so than the suppression caused by the tumor would be the reason for the lack of antibodies
If this immune suppression could be overcome in vitro it could lead to the formation of antibodies that are not detectable in the serum We call this process Cimmunology Identifying the structures that the antibodies bind to and looking for a correlation between the tumor pathology and the antibodies specificity - could provide additional information about the tumor - via a blood test
12 Future CAAb Test
The Cimmunology based test is comprised of an in-vitro stimulation step and an antibody detection step
The assay will be used to find cancer associated antibodies in Breast cancer patients
Identification of Breast cancer associated antibodies could provide the clinician with additional immunological and antigenic information regarding the patients condition or medical status
The Cimmunology process is comprised of a blood collection and processing step and an incubation in a special stimulation tube called CimTube period in a humid 37oC incubator with 5 CO2
The test for the detection of specific antibodies is ELISA based and includes different potentially interesting antigens
The relevant medical and pathological information to be collected form the patients file will enable a more detailed analysis of the test results
0 STUDY OBJECTIVES
21 Primary objective
To find Cancer Associated Antibodies
22 Effectiveness
To determine the ability of the Cimmunology process to lead to in vitro antibody production the ability of the ELISA assays to detect any of those antibodies and to establish the relationship between the ELISA results and the clinical pathological status of the patient The statistical significance of the CAAb test results will be determined
30 STUDY POPULATIONS AND PATIENT SELECTION
31 Study Population
311 Breast cancer patients The study population will include subjects that have been diagnosed with suspected breast cancer ie positive biopsy prior to any surgical procedure or any other anti-cancer treatment
312 Control group
The control group will include patients at breast cancer centers or any other appropriate care unit with negative Mammography in the last 3 months
32 Participating Centers
321 General - The study will be based on multi-centers participation
322 Locations - All sites will be within Hospitals either in the Breast Cancer centers or in the relevant specific unit which is responsible for providing care for the BC patients The minimum number of sites for the study is 2 and the max is 10
STUDY DESIGN
41 Overall Description
The current study is a comparative observational two-arm study involving all consecutive Breast cancer patients during the study duration and age matched control subjects The purpose of the study is to assess the effectiveness of the CAAb test in detecting Breast cancer associated antibodies
Subjects will be screened for potential participation in the study according to the inclusion and exclusion criteria Patients recruited will be asked to sign an informed consent form
The data on the patients will include parameters of the clinical and pathological state of the patients results of CimTube culture step and the experimental kits The effectiveness of the CAAb test will be assessed by evaluating the statistical significance of the CAAb test used
The study will utilize a three step group-sequential design with two interim and one final analysis using the OBrian-Fleming boundaries The data collection may be stopped after the first or second interim analysis if the bounds are reached This could substantially decrease the number of patients and the time of the study while just slightly increasing the maximum number of patients See Sample size considerations below
We will use 12 ratio between cancer and control patients to decrease the necessary number of cancer patients which is the main limiting factor
The sponsor will conduct interim analysis following the first third and second third of Breast cancer cases and the sample size will be recalculated accordingly to the obtained estimates of the mean and variance of the test in the two groups
Therefore at least 100 patients will be enrolled in the study The control group will be of 200 patients
42 Study Procedure
In the hospital
First an identified study patient has to sign an informed consent form and her Eligibility Form filled see appendix B both should be bar-coded
Patient demographic and clinical information acquired from the patients medical file including age country of origin medical history and results of tests done leading to the diagnostic evaluation of Breast cancer not relevant in the control group will be recorded on the appropriate bar-coded pre-study case report forms see appendix C CRF-BC and CRF-BC-Control The reports will be either electronic a dedicated and secured internet site or via a hard copy A hard copy of the records will be kept in the department The study is anonymous and the Department will keep the name of the patient without reveling it to investigators and to the study sponsor
The Study Sponsor will provide the tubes and the barcode labels for monitoring The doctor nurse phlebotomist will collect three heparin vacuum tubes 20-24 ml label bar-code them and fill up the initial step in the Sample follow-up form SFF see appendix D The tubes with the blood will be packaged in double sealed containers will be provided by the sponsor The department will notify the designated shipper to collect the blood up to 1 hour from the time that it has been sampled
For each BC sample two age matched controls - 3 years will be recruited too
Transportation from the Hospital to the Handling Lab
The blood will be transported at room temperature 18-250C according to the relevant regulations to the laboratory Transportation time will be not more than 6 hours Times to be recorded in SFF
In the blood handling laboratory
The blood handling laboratory to be located no further than 200km from the collection site will isolate PBMC Peripheral blood mononuclear cells and put them into Cimmunology culture within 20 hours form the sampling time fill up time in SFF
The detailed protocol and the Cimmunology media will be provided by Lab Discoveries to the participating laboratories where applicable All hospitals in Israel will send the blood samples to the laboratory at Lab Discoveries After the culture step the culture fluid will be collected and frozen at -80 in properly labeled aliquots The frozen samples will be sent to the diagnostic laboratory for antibody tests
In the diagnostic laboratory
Antibody tests such as ELISA will be prepared using different antigens The samples will be tested for response to the two antigens that have been identified in the past and for additional new ones All the data will be permanently recorded directly into a dedicated computer For each sample all antibody results will be put into the coded patient file The results will be analyzed by a statistician with expertise in cancer population studies
At the Hospital post surgery
2-3 weeks after the surgery the CRO will collect from the patients file the results from the pathology lab and any other relevant information as to the nature and state of the tumor that was removed during surgery These results will be recorded on the dedicated and secured internet site and a printed bar-coded CRFs will be kept in the Department
90 STATISTICAL CONSIDERATIONS
The present study is a comparative observational two-arm study involving all consecutive Breast cancer patients during the study duration and age matched control subjects The main purpose of the study is to assess the effectiveness of the CAAb tests This effectiveness will be measured by the Fisher distance often called effect size d defined as
Where m1 is the mean value of the test in the controls m2 is the mean value in the Breast cancer patients m1-m2 denotes the absolute value of the difference m1-m2 and s is the common estimate of the standard deviation of the distribution of the test in one group control or Breast cancer
91 Statistical Hypothesis
Null Hypothesis There is no relationship between the presence or absence of BC and the CAAb ie d0
Alternative hypothesis The expectation of the CAAb in the cancer population differ from that of the control population ie m1 not equal to m2 Since the sign of the difference is not important the test will be two-sided
92 General Considerations
According to our previous results we assume that
1 The distributions of the test results in each group are close to Normal or may be transformed to the Normal by log-transformation
2 The variance of the distributions in the two groups are approximately equal
3 Having in mind that this is a pilot study we will consider the results as significant if the two-sided p-value is 01 or less The bound 01 instead of 005 was chosen to decrease the chances of random exclusion of some potentially informative tests
When deciding the continuation of the study no correction for multiple comparisons is assumed However the results will be presented without and with the correction using the FDR of BenjaminiHochberg approach Benjamini Hochberg 1995
Where confidence limits are appropriate the confidence level will be 95
93 Sample Size The sample size was calculated using relevant information from our previous study on Peptides P3 and P1 and Breast cancer There is no information in the literature regarding cancer associated antibodies
The sample size is calculated for a three step sequential group design with two sided significance 01 and 005 power 08 for cancer control ratio 12 for D of 05 The calculations used the standard formulas for sample size necessary for comparison mean values in two independent groups by t-test The correction for a group-sequential procedure with OBien and Flemings test increases the numbers by 1017 and 1027 for significance values 005 and 01 that is negligible for our sample sizes see C Jennison BWTurnbill Group Sequential Methods with Applications to Clinical Trials ChapmannHall2000 Boca Raton FL US pp30 table 24
The calculations were done using NCSS-PASS software
94 Endpoints Evaluation
941 Effectiveness Analysis
A calculated D between 03 and 05 will mark a test as non effective to be used alone However it may be important in combination with other tests as a part of the multivariate test procedure Tests with d03 will be considered as non-effective A test or a combination of tests with d05 will mark the CAAb test as effective
Antibodies are a specific response to any foreign antigen and they are usually detectable in the serum within 5-7 days after the initial exposure to it However there are some cases where there is a suppression of the specific immune response such as in the case of a tumor To date there are no reports on serum antibodies that are associated to cancer Some tumors when surgically removed and studied have been found to have both infiltrates of T lymphocytes and antibodies bound to the tumor cells These antibodies and other specific immune responses are assumed to be formed at later stages of the tumor growth however even in those cases serum antibodies have not been reported probably due to them being mostly absorbed by the tumor mass
Since the mutations of a normal cell that lead to a malignant cell mass involves changes in the structure and function of the cells it could be assumed that the immune system has recognized some of these changes as antigenic determinates that should be responded against if so than the suppression caused by the tumor would be the reason for the lack of antibodies
If this immune suppression could be overcome in vitro it could lead to the formation of antibodies that are not detectable in the serum We call this process Cimmunology Identifying the structures that the antibodies bind to and looking for a correlation between the tumor pathology and the antibodies specificity - could provide additional information about the tumor - via a blood test
12 Future CAAb Test
The Cimmunology based test is comprised of an in-vitro stimulation step and an antibody detection step
The assay will be used to find cancer associated antibodies in Breast cancer patients
Identification of Breast cancer associated antibodies could provide the clinician with additional immunological and antigenic information regarding the patients condition or medical status
The Cimmunology process is comprised of a blood collection and processing step and an incubation in a special stimulation tube called CimTube period in a humid 37oC incubator with 5 CO2
The test for the detection of specific antibodies is ELISA based and includes different potentially interesting antigens
The relevant medical and pathological information to be collected form the patients file will enable a more detailed analysis of the test results
0 STUDY OBJECTIVES
21 Primary objective
To find Cancer Associated Antibodies
22 Effectiveness
To determine the ability of the Cimmunology process to lead to in vitro antibody production the ability of the ELISA assays to detect any of those antibodies and to establish the relationship between the ELISA results and the clinical pathological status of the patient The statistical significance of the CAAb test results will be determined
30 STUDY POPULATIONS AND PATIENT SELECTION
31 Study Population
311 Breast cancer patients The study population will include subjects that have been diagnosed with suspected breast cancer ie positive biopsy prior to any surgical procedure or any other anti-cancer treatment
312 Control group
The control group will include patients at breast cancer centers or any other appropriate care unit with negative Mammography in the last 3 months
32 Participating Centers
321 General - The study will be based on multi-centers participation
322 Locations - All sites will be within Hospitals either in the Breast Cancer centers or in the relevant specific unit which is responsible for providing care for the BC patients The minimum number of sites for the study is 2 and the max is 10
STUDY DESIGN
41 Overall Description
The current study is a comparative observational two-arm study involving all consecutive Breast cancer patients during the study duration and age matched control subjects The purpose of the study is to assess the effectiveness of the CAAb test in detecting Breast cancer associated antibodies
Subjects will be screened for potential participation in the study according to the inclusion and exclusion criteria Patients recruited will be asked to sign an informed consent form
The data on the patients will include parameters of the clinical and pathological state of the patients results of CimTube culture step and the experimental kits The effectiveness of the CAAb test will be assessed by evaluating the statistical significance of the CAAb test used
The study will utilize a three step group-sequential design with two interim and one final analysis using the OBrian-Fleming boundaries The data collection may be stopped after the first or second interim analysis if the bounds are reached This could substantially decrease the number of patients and the time of the study while just slightly increasing the maximum number of patients See Sample size considerations below
We will use 12 ratio between cancer and control patients to decrease the necessary number of cancer patients which is the main limiting factor
The sponsor will conduct interim analysis following the first third and second third of Breast cancer cases and the sample size will be recalculated accordingly to the obtained estimates of the mean and variance of the test in the two groups
Therefore at least 100 patients will be enrolled in the study The control group will be of 200 patients
42 Study Procedure
In the hospital
First an identified study patient has to sign an informed consent form and her Eligibility Form filled see appendix B both should be bar-coded
Patient demographic and clinical information acquired from the patients medical file including age country of origin medical history and results of tests done leading to the diagnostic evaluation of Breast cancer not relevant in the control group will be recorded on the appropriate bar-coded pre-study case report forms see appendix C CRF-BC and CRF-BC-Control The reports will be either electronic a dedicated and secured internet site or via a hard copy A hard copy of the records will be kept in the department The study is anonymous and the Department will keep the name of the patient without reveling it to investigators and to the study sponsor
The Study Sponsor will provide the tubes and the barcode labels for monitoring The doctor nurse phlebotomist will collect three heparin vacuum tubes 20-24 ml label bar-code them and fill up the initial step in the Sample follow-up form SFF see appendix D The tubes with the blood will be packaged in double sealed containers will be provided by the sponsor The department will notify the designated shipper to collect the blood up to 1 hour from the time that it has been sampled
For each BC sample two age matched controls - 3 years will be recruited too
Transportation from the Hospital to the Handling Lab
The blood will be transported at room temperature 18-250C according to the relevant regulations to the laboratory Transportation time will be not more than 6 hours Times to be recorded in SFF
In the blood handling laboratory
The blood handling laboratory to be located no further than 200km from the collection site will isolate PBMC Peripheral blood mononuclear cells and put them into Cimmunology culture within 20 hours form the sampling time fill up time in SFF
The detailed protocol and the Cimmunology media will be provided by Lab Discoveries to the participating laboratories where applicable All hospitals in Israel will send the blood samples to the laboratory at Lab Discoveries After the culture step the culture fluid will be collected and frozen at -80 in properly labeled aliquots The frozen samples will be sent to the diagnostic laboratory for antibody tests
In the diagnostic laboratory
Antibody tests such as ELISA will be prepared using different antigens The samples will be tested for response to the two antigens that have been identified in the past and for additional new ones All the data will be permanently recorded directly into a dedicated computer For each sample all antibody results will be put into the coded patient file The results will be analyzed by a statistician with expertise in cancer population studies
At the Hospital post surgery
2-3 weeks after the surgery the CRO will collect from the patients file the results from the pathology lab and any other relevant information as to the nature and state of the tumor that was removed during surgery These results will be recorded on the dedicated and secured internet site and a printed bar-coded CRFs will be kept in the Department
90 STATISTICAL CONSIDERATIONS
The present study is a comparative observational two-arm study involving all consecutive Breast cancer patients during the study duration and age matched control subjects The main purpose of the study is to assess the effectiveness of the CAAb tests This effectiveness will be measured by the Fisher distance often called effect size d defined as
Where m1 is the mean value of the test in the controls m2 is the mean value in the Breast cancer patients m1-m2 denotes the absolute value of the difference m1-m2 and s is the common estimate of the standard deviation of the distribution of the test in one group control or Breast cancer
91 Statistical Hypothesis
Null Hypothesis There is no relationship between the presence or absence of BC and the CAAb ie d0
Alternative hypothesis The expectation of the CAAb in the cancer population differ from that of the control population ie m1 not equal to m2 Since the sign of the difference is not important the test will be two-sided
92 General Considerations
According to our previous results we assume that
1 The distributions of the test results in each group are close to Normal or may be transformed to the Normal by log-transformation
2 The variance of the distributions in the two groups are approximately equal
3 Having in mind that this is a pilot study we will consider the results as significant if the two-sided p-value is 01 or less The bound 01 instead of 005 was chosen to decrease the chances of random exclusion of some potentially informative tests
When deciding the continuation of the study no correction for multiple comparisons is assumed However the results will be presented without and with the correction using the FDR of BenjaminiHochberg approach Benjamini Hochberg 1995
Where confidence limits are appropriate the confidence level will be 95
93 Sample Size The sample size was calculated using relevant information from our previous study on Peptides P3 and P1 and Breast cancer There is no information in the literature regarding cancer associated antibodies
The sample size is calculated for a three step sequential group design with two sided significance 01 and 005 power 08 for cancer control ratio 12 for D of 05 The calculations used the standard formulas for sample size necessary for comparison mean values in two independent groups by t-test The correction for a group-sequential procedure with OBien and Flemings test increases the numbers by 1017 and 1027 for significance values 005 and 01 that is negligible for our sample sizes see C Jennison BWTurnbill Group Sequential Methods with Applications to Clinical Trials ChapmannHall2000 Boca Raton FL US pp30 table 24
The calculations were done using NCSS-PASS software
94 Endpoints Evaluation
941 Effectiveness Analysis
A calculated D between 03 and 05 will mark a test as non effective to be used alone However it may be important in combination with other tests as a part of the multivariate test procedure Tests with d03 will be considered as non-effective A test or a combination of tests with d05 will mark the CAAb test as effective
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None