<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT01688232</url>
  </required_header>
  <id_info>
    <org_study_id>NI11033</org_study_id>
    <nct_id>NCT01688232</nct_id>
  </id_info>
  <brief_title>The Immunoscore as a Prognostic Marker for Patients With a Colorectal Cancer</brief_title>
  <acronym>IMMUCOL</acronym>
  <official_title>Colorectal Cancer: a Prospective Multicentric Study of the in Situ Immune Infiltrate for the Identification of the Patients at High Risk of Relapse</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Assistance Publique - Hôpitaux de Paris</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Ministry of Health, France</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Assistance Publique - Hôpitaux de Paris</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Medical and Scientific Background: Colorectal cancers (CRC) are the leading cancers in both&#xD;
      genders. This cancer is of prognostic severity. The prognosis of this cancer has not been&#xD;
      significantly improved. The treatment of colonic cancer is primarily based on surgery.&#xD;
      Adjuvant chemotherapy is proposed for metastatic cancers (stages III and IV). However nearly&#xD;
      30% of patients with localized cancer (stage II) will present a recurrence. Despite intense&#xD;
      research efforts, no markers of sufficient prognostic value (independent of TNM) are&#xD;
      available to identify this group of patients and justify intensified therapy. The natural&#xD;
      history of cancer involves interactions between the tumor and the immune system of the host.&#xD;
      The immune infiltration at the tumor site may be indicative of host response. We showed that&#xD;
      the density of intratumor memory T cells (CD45RO) and cytotoxic T cells (CD8) in tumor&#xD;
      regions (the core and the invasive margin) influenced the occurrence of early events of&#xD;
      metastasis (tumor emboli) and was strongly associated with prognosis of patients with CRC&#xD;
      (Pagès et al, NEJM-2005). This immune criterion was a better predictor of survival than the&#xD;
      &quot;gold standard&quot; histoprognostic data of tumor invasion (T stage and N) and allowed to&#xD;
      identify a group of patients at high risk of recurrence (*2). The availability of this immune&#xD;
      criterion in clinical practice could improve the prognostic assessment of patients and better&#xD;
      guide the therapeutic. A dedicated platform has been implemented in our hospital to speed up&#xD;
      the transfer of this immune investigation into the clinic. The investigation takes into&#xD;
      account the density of immune cell populations in tumor regions (the core and the invasive&#xD;
      front of the tumor). This methodology has been validated for the markers CD45RO, CD3 and CD8&#xD;
      and leads to the creation of an immune score (&quot;immunoscore&quot; ranging from IO to I4). We&#xD;
      validated the prognostic impact of the immunoscore in a retrospective series of 250&#xD;
      colorectal cancers. The main objective of this multicentric prospective clinical study is to&#xD;
      assess in clinical practice the &quot;immunoscore&quot; and measure its prognostic value. The cohort&#xD;
      study will include 400 patients with CRC stage I to IV (6 centers for inclusion; Paris-HEGP,&#xD;
      Dijon, Bobigny-Avicenne, Besancon, Poitiers, Rouen). Two years of inclusion and a follow-up&#xD;
      of 3 years for each patient will be performed (co-financing for data collection during the&#xD;
      4th and 5th year planned). For each patient, the pathologist of the center will send to the&#xD;
      immunomonitoring platform of HEGP tissue sections from a paraffin block containing the tumor&#xD;
      regions. The investigations will combine a step of immunostaining for CD3, CD8, and CD45RO&#xD;
      (Ventana automate) markers, high-resolution scanning of the stained slides and quantification&#xD;
      of digital images by an imaging module developed by our group from the program of Developer&#xD;
      XD of Définiens company. The immunoscore will be calculated and the score will be correlated&#xD;
      to the clinical data for the relapse and the survival. The secondary objective of this&#xD;
      program is to evaluate the prognostic performance of the immune infiltrate on the biopsies&#xD;
      performed for diagnostic purposes. This study will be conducted on patients of the cohort&#xD;
      whose biopsies were performed in the same hospital than surgery and whose samples are&#xD;
      available in pathology laboratories involved (representing 50% of cases). For the same&#xD;
      patients will be conducted a genetic investigation of the tumor to assess the MSI status, the&#xD;
      presence of a K-Ras and BRAF mutations. This investigation will be performed on tumor&#xD;
      sections from the same tumor block selected for the immunohistochemical analyses. The&#xD;
      sections will be processed by the Department of Biology, Hôpital Européen Georges Pompidou.&#xD;
      The prognostic performance of the immune investigations performed on tumor sections and&#xD;
      biopsies will be compared to that of genetic features of the tumor. Finally, a questionnaire&#xD;
      will be sent every six months along the monitoring of the patients to obtain information&#xD;
      concerning (i) the emergence of an immune disorder (such as allergy, autoimmunity,&#xD;
      inflammatory process) and (ii) the psychological status of the patients. The potential impact&#xD;
      of such parameters during the course of the disease on the prediction of the relapse and&#xD;
      survival obtained with the immunoscore performed at the time of surgery will be evaluated.&#xD;
      Expected results : This prospective study is an indispensable step for the clinical&#xD;
      validation of the prognostic value of the immunoscore. The secondary objectives should help&#xD;
      to precise the benefit of a concomitant analysis of the biopsy, the genetic features of the&#xD;
      tumor. The questionnaire should help to identify the clinical parameters to track along the&#xD;
      monitoring&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      The main objective is to assess the impact of the immunoscore in the clinical practice. To&#xD;
      this end, the immune parameter (CD3, CD8, CD45RO) of colorectal cancers (stage I-IV) will be&#xD;
      evaluated prospectively through a large multicenter study. 400 patients with CRC stage I to&#xD;
      IV will be included (6 centers for inclusion; Paris-HEGP, Dijon, Bobigny-Avicenne, Besancon,&#xD;
      Poitiers, Rouen). The number of patients to be included has been calculated on the basis of&#xD;
      an inclusion period of 24 months, a follow-up of 3 years (a collection of data during the 4th&#xD;
      and 5th year is planned with a co-financing), an expected event rate of 20%, a prognostic&#xD;
      value of the immune parameter similar to that observed in the retrospective study of 399&#xD;
      patients with CRC (*1) and a power of the test of 90%. During both initial management and&#xD;
      follow-up, all therapeutic procedures will be registered (surgery, radiotherapy,&#xD;
      chemotherapy…). Data will be collected from clinical examination, radiological exams and&#xD;
      biological tests that are usually performed during a hospitalization for colorectal cancer:&#xD;
      Full clinical examination and questioning about the presence of personal and family history&#xD;
      of CRC, adenomas and other cancers; Abdominal ultrasound, chest X-ray; Computed tomography&#xD;
      (CT) and magnetic resonance imaging (MRI) according to clinician's requirement; Standard&#xD;
      laboratory tests and blood assay for carcinoembryonic antigen (CEA); The Histopathologic&#xD;
      study of the tumor containing the status of the surgical excision margins, the histological&#xD;
      type of cancer, the level of parietal invasion, the number of lymph nodes examined and the&#xD;
      number of metastatic lymph nodes, according to the TNM classification. Patients will be&#xD;
      followed-up every 3 months during the first 2 years, and every 6 months for the rest of the&#xD;
      follow-up period, as recommended. Each patient will be followed up to 5 years. Clinical,&#xD;
      biological and radiological data will be registered into a specific CRF. Verification of&#xD;
      inclusions and monitoring of data will be conducted by the HEGP Clinical Research Unit (URC&#xD;
      HEGP) and the principal investigator. A plan for data-management will be built in line with&#xD;
      the monitoring. A questionnaire will also be sent to the patients every six month for five&#xD;
      years. This questionnaire was built by Pr F Pagès and by psychologists and psychiatrists to&#xD;
      search for information concerning (i) the presence of an immune disorder (such as allergy,&#xD;
      autoimmunity, inflammatory process) and (ii) the psychological status of the patients (eg.&#xD;
      depression). After monitoring, all data will be stored using a double data entry system in a&#xD;
      MS-Access database designed by the URC-HEGP and in the TME.dB database created by the team of&#xD;
      the principal investigator. The 2 databases will be merged, and the final data base will be&#xD;
      frozen for statistical analysis. Sample preparation: The pathologist of each center will&#xD;
      choose a tumor block containing the core of the tumor and the invasive margin. This tumor&#xD;
      block is part of the tumor samples routinely processed by the department of Pathology for&#xD;
      diagnosis. three tissue sections of 4-microns will be made and deposit on a glass slide to&#xD;
      perform the immunohistochemistry. Four tissue sections of 4-microns will be made and deposit&#xD;
      on an Eppendorf safe lock tubes to perform the genetic analyses. Anonymous patient names on&#xD;
      the slides and tubes will be provided by using a code number and initials of the patient. If&#xD;
      biopsies for diagnostic purposes are available (i.e. those performed for the patients during&#xD;
      their initial hospitalization (50% of cases), A tissue sections of each biopsy will be done.&#xD;
      All slides and tubes will be sent by mail to the HEGP platform for analysis of the immune&#xD;
      infiltrate and to the Biology department for the genetic analyses. The immunoscore will be&#xD;
      determine using the following steps: Immunostainings for CD3, CD8, CD45RO on Benchmark XT&#xD;
      automate; Scanning the immunostainings on Hamamatsu scanner: Quantification of the immune&#xD;
      populations in specific tumor areas (the core and the invasive margin) with a dedicated image&#xD;
      analysis module based on the recognition of histological structures adapted to colorectal&#xD;
      cancer. This module has been developed by our group in connection with the Definiens company.&#xD;
      A semi-automatic procedure allows the quantification of stained cells in each tumor region.&#xD;
      Each region is divided into tiles of 800 microns side (500 to 700 tiles analyzed per tumor).&#xD;
      The density of each marker is calculated using the mean density of the three tiles the most&#xD;
      infiltrated in each tumor region. A categorization into high density (Hi) or low (Lo) for&#xD;
      each marker in each tumor region of interest will be performed. This categorization is done&#xD;
      by comparing the observed density with the optimal cut-off density (min p value approach) we&#xD;
      previously determined for the DFS and OS in a retrospective study of 250 CRC. Genetic&#xD;
      analyses: The tubes containing the sections for the genetic analyses of patients with inform&#xD;
      consent signed for genetic investigation will be transfer to the Department of Biology,&#xD;
      Hôpital Européen Georges Pompidou. The % of tumor cells of each specimen will be determined&#xD;
      by a pathologist (based on a H&amp;E evaluation; less than 10%, 10-20%, 20-50%, up to 50% tumor&#xD;
      cells) and the DNA extraction will be performed. Genetic investigation will comprise the MSI&#xD;
      status, the presence of a K-Ras and BRAF mutation&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Active, not recruiting</overall_status>
  <start_date type="Actual">September 27, 2012</start_date>
  <completion_date type="Anticipated">December 31, 2021</completion_date>
  <primary_completion_date type="Anticipated">December 31, 2021</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Cohort</observational_model>
    <time_perspective>Prospective</time_perspective>
  </study_design_info>
  <primary_outcome>
    <measure>relapse in relation with the immunoscore determined on a tumor section.</measure>
    <time_frame>evry 3 month during 3 years and every six months during the fourth year</time_frame>
    <description>this primary outcome (relapse) will be correlated to the immunoscore determined in the primary tumor (eg. the density of immune cells in tumor regions)</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>relapse in relation with the immunoscore on biopsies</measure>
    <time_frame>evry 3 month during 3 years and every six months during the fourth year</time_frame>
    <description>this primary outcome (relapse) will be correlated to the immunoscore determined in biopsies performed for diagnosis purpose (eg. the density of immune cells in the biopsies specimens)</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>relapse in relation with the genetic features of the tumor</measure>
    <time_frame>evry 3 month during 3 years and every six months during the fourth year</time_frame>
    <description>this primary outcome (relapse) will be correlated to the genetic features of the tumor (eg. MSI status, K-Ras and BRAF mutations). The prognostic performance of the genetic and immunologic analyses will be compared.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>relapse in relation with the immunological events and psychological status of the patient during the monitoring</measure>
    <time_frame>every six months during 4 years</time_frame>
    <description>a questionnaire will be sent every six months along the monitoring of the patients to obtain informations concerning (i) the emergence of an immune disorder and (ii) the psychological status of the patients. The potential impact of such parameters during the course of the disease on the prediction of the relapse and survival obtained with the immunoscore performed at the time of surgery will be evaluated. The questionnaire should help to identify the clinical parameters to track along the monitoring.</description>
  </secondary_outcome>
  <enrollment type="Actual">459</enrollment>
  <condition>Colorectal Cancer</condition>
  <condition>Surgical Resection of the Colorectal Tumor</condition>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      The pathologist of each center will choose a tumor block containing the core of the tumor and&#xD;
      the invasive margin. Three tissue sections of 4 microns of this tumor block will be cut and&#xD;
      deposited on glass slides for the immunohistochemical analyses. Four tissue sections of 4&#xD;
      microns will be cut and deposited in a safe lock eppendorf tube for a DNA extraction. If&#xD;
      biopsies for diagnostic purpose are available in the laboratory of pathology for the same&#xD;
      patient, one tissue section of 4 microns of each biopsy will be cut and deposited on a glass&#xD;
      slide&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Adult patient with newly diagnosed colorectal cancer.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Non-Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Adult patient with newly diagnosed colorectal cancer.&#xD;
&#xD;
          -  Patient with signed informed consent.&#xD;
&#xD;
          -  Follow up made by the clinical center for inclusion or by a medical team in relation&#xD;
             with the center.&#xD;
&#xD;
        Exclusion Criteria:&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>N/A</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Franck Pagès, Professor (MD-PHD)</last_name>
    <role>Principal Investigator</role>
    <affiliation>AP-HP; Paris Descartes University</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Hopital de Besançon (CHU)</name>
      <address>
        <city>Besançon</city>
        <zip>25000</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Hopital Avicenne (CHU)</name>
      <address>
        <city>Bobigny</city>
        <zip>93000</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Hopital de Dijon (CHU)</name>
      <address>
        <city>Dijon</city>
        <zip>21000</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Hopital Europeen Georges Pompidou (HEGP)</name>
      <address>
        <city>Paris</city>
        <zip>75015</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Hopital de Poitiers (CHU)</name>
      <address>
        <city>Poitiers</city>
        <zip>86000</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location>
    <facility>
      <name>Hopital Charles Nicolle(CHU)</name>
      <address>
        <city>Rouen</city>
        <zip>76000</zip>
        <country>France</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>France</country>
  </location_countries>
  <results_reference>
    <citation>Galon J, Costes A, Sanchez-Cabo F, Kirilovsky A, Mlecnik B, Lagorce-Pages C, Tosolini M, Camus M, Berger A, Wind P, Zinzindohoue F, Bruneval P, Cugnenc PH, Trajanoski Z, Fridman WH, Pages F. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science. 2006 Sep 29;313(5795):1960-4. doi: 10.1126/science.1129139.</citation>
    <PMID>17008531</PMID>
  </results_reference>
  <results_reference>
    <citation>Pages F, Kirilovsky A, Mlecnik B, Asslaber M, Tosolini M, Bindea G, Lagorce C, Wind P, Marliot F, Bruneval P, Zatloukal K, Trajanoski Z, Berger A, Fridman WH, Galon J. In situ cytotoxic and memory T cells predict outcome in patients with early-stage colorectal cancer. J Clin Oncol. 2009 Dec 10;27(35):5944-51. doi: 10.1200/JCO.2008.19.6147. Epub 2009 Oct 26.</citation>
    <PMID>19858404</PMID>
  </results_reference>
  <results_reference>
    <citation>Mlecnik B, Tosolini M, Kirilovsky A, Berger A, Bindea G, Meatchi T, Bruneval P, Trajanoski Z, Fridman WH, Pages F, Galon J. Histopathologic-based prognostic factors of colorectal cancers are associated with the state of the local immune reaction. J Clin Oncol. 2011 Feb 20;29(6):610-8. doi: 10.1200/JCO.2010.30.5425. Epub 2011 Jan 18.</citation>
    <PMID>21245428</PMID>
  </results_reference>
  <verification_date>October 2021</verification_date>
  <study_first_submitted>September 14, 2012</study_first_submitted>
  <study_first_submitted_qc>September 14, 2012</study_first_submitted_qc>
  <study_first_posted type="Estimate">September 19, 2012</study_first_posted>
  <last_update_submitted>October 11, 2021</last_update_submitted>
  <last_update_submitted_qc>October 11, 2021</last_update_submitted_qc>
  <last_update_posted type="Actual">October 12, 2021</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <keyword>colorectal cancer</keyword>
  <keyword>tumor resection</keyword>
  <keyword>immunoscore</keyword>
  <keyword>lymphocytic density</keyword>
  <keyword>CD3</keyword>
  <keyword>CD8</keyword>
  <keyword>CD45RO</keyword>
  <keyword>biopsy</keyword>
  <keyword>genetic feature of the tumor</keyword>
  <keyword>relapse</keyword>
  <keyword>immunohistochemistry</keyword>
  <keyword>image analysis software</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Colorectal Neoplasms</mesh_term>
  </condition_browse>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT01688232
org study id: NI11033
nct id: NCT01688232
lead sponsor: 
collaborator: 
has dmc: No
Medical and Scientific Background: Colorectal cancers (CRC) are the leading cancers in both
      genders. This cancer is of prognostic severity. The prognosis of this cancer has not been
      significantly improved. The treatment of colonic cancer is primarily based on surgery.
      Adjuvant chemotherapy is proposed for metastatic cancers (stages III and IV). However nearly
      30% of patients with localized cancer (stage II) will present a recurrence. Despite intense
      research efforts, no markers of sufficient prognostic value (independent of TNM) are
      available to identify this group of patients and justify intensified therapy. The natural
      history of cancer involves interactions between the tumor and the immune system of the host.
      The immune infiltration at the tumor site may be indicative of host response. We showed that
      the density of intratumor memory T cells (CD45RO) and cytotoxic T cells (CD8) in tumor
      regions (the core and the invasive margin) influenced the occurrence of early events of
      metastasis (tumor emboli) and was strongly associated with prognosis of patients with CRC
      (Pagès et al, NEJM-2005). This immune criterion was a better predictor of survival than the
      "gold standard" histoprognostic data of tumor invasion (T stage and N) and allowed to
      identify a group of patients at high risk of recurrence (*2). The availability of this immune
      criterion in clinical practice could improve the prognostic assessment of patients and better
      guide the therapeutic. A dedicated platform has been implemented in our hospital to speed up
      the transfer of this immune investigation into the clinic. The investigation takes into
      account the density of immune cell populations in tumor regions (the core and the invasive
      front of the tumor). This methodology has been validated for the markers CD45RO, CD3 and CD8
      and leads to the creation of an immune score ("immunoscore" ranging from IO to I4). We
      validated the prognostic impact of the immunoscore in a retrospective series of 250
      colorectal cancers. The main objective of this multicentric prospective clinical study is to
      assess in clinical practice the "immunoscore" and measure its prognostic value. The cohort
      study will include 400 patients with CRC stage I to IV (6 centers for inclusion; Paris-HEGP,
      Dijon, Bobigny-Avicenne, Besancon, Poitiers, Rouen). Two years of inclusion and a follow-up
      of 3 years for each patient will be performed (co-financing for data collection during the
      4th and 5th year planned). For each patient, the pathologist of the center will send to the
      immunomonitoring platform of HEGP tissue sections from a paraffin block containing the tumor
      regions. The investigations will combine a step of immunostaining for CD3, CD8, and CD45RO
      (Ventana automate) markers, high-resolution scanning of the stained slides and quantification
      of digital images by an imaging module developed by our group from the program of Developer
      XD of Définiens company. The immunoscore will be calculated and the score will be correlated
      to the clinical data for the relapse and the survival. The secondary objective of this
      program is to evaluate the prognostic performance of the immune infiltrate on the biopsies
      performed for diagnostic purposes. This study will be conducted on patients of the cohort
      whose biopsies were performed in the same hospital than surgery and whose samples are
      available in pathology laboratories involved (representing 50% of cases). For the same
      patients will be conducted a genetic investigation of the tumor to assess the MSI status, the
      presence of a K-Ras and BRAF mutations. This investigation will be performed on tumor
      sections from the same tumor block selected for the immunohistochemical analyses. The
      sections will be processed by the Department of Biology, Hôpital Européen Georges Pompidou.
      The prognostic performance of the immune investigations performed on tumor sections and
      biopsies will be compared to that of genetic features of the tumor. Finally, a questionnaire
      will be sent every six months along the monitoring of the patients to obtain information
      concerning (i) the emergence of an immune disorder (such as allergy, autoimmunity,
      inflammatory process) and (ii) the psychological status of the patients. The potential impact
      of such parameters during the course of the disease on the prediction of the relapse and
      survival obtained with the immunoscore performed at the time of surgery will be evaluated.
      Expected results : This prospective study is an indispensable step for the clinical
      validation of the prognostic value of the immunoscore. The secondary objectives should help
      to precise the benefit of a concomitant analysis of the biopsy, the genetic features of the
      tumor. The questionnaire should help to identify the clinical parameters to track along the
      monitoring
The main objective is to assess the impact of the immunoscore in the clinical practice. To
      this end, the immune parameter (CD3, CD8, CD45RO) of colorectal cancers (stage I-IV) will be
      evaluated prospectively through a large multicenter study. 400 patients with CRC stage I to
      IV will be included (6 centers for inclusion; Paris-HEGP, Dijon, Bobigny-Avicenne, Besancon,
      Poitiers, Rouen). The number of patients to be included has been calculated on the basis of
      an inclusion period of 24 months, a follow-up of 3 years (a collection of data during the 4th
      and 5th year is planned with a co-financing), an expected event rate of 20%, a prognostic
      value of the immune parameter similar to that observed in the retrospective study of 399
      patients with CRC (*1) and a power of the test of 90%. During both initial management and
      follow-up, all therapeutic procedures will be registered (surgery, radiotherapy,
      chemotherapy…). Data will be collected from clinical examination, radiological exams and
      biological tests that are usually performed during a hospitalization for colorectal cancer:
      Full clinical examination and questioning about the presence of personal and family history
      of CRC, adenomas and other cancers; Abdominal ultrasound, chest X-ray; Computed tomography
      (CT) and magnetic resonance imaging (MRI) according to clinician's requirement; Standard
      laboratory tests and blood assay for carcinoembryonic antigen (CEA); The Histopathologic
      study of the tumor containing the status of the surgical excision margins, the histological
      type of cancer, the level of parietal invasion, the number of lymph nodes examined and the
      number of metastatic lymph nodes, according to the TNM classification. Patients will be
      followed-up every 3 months during the first 2 years, and every 6 months for the rest of the
      follow-up period, as recommended. Each patient will be followed up to 5 years. Clinical,
      biological and radiological data will be registered into a specific CRF. Verification of
      inclusions and monitoring of data will be conducted by the HEGP Clinical Research Unit (URC
      HEGP) and the principal investigator. A plan for data-management will be built in line with
      the monitoring. A questionnaire will also be sent to the patients every six month for five
      years. This questionnaire was built by Pr F Pagès and by psychologists and psychiatrists to
      search for information concerning (i) the presence of an immune disorder (such as allergy,
      autoimmunity, inflammatory process) and (ii) the psychological status of the patients (eg.
      depression). After monitoring, all data will be stored using a double data entry system in a
      MS-Access database designed by the URC-HEGP and in the TME.dB database created by the team of
      the principal investigator. The 2 databases will be merged, and the final data base will be
      frozen for statistical analysis. Sample preparation: The pathologist of each center will
      choose a tumor block containing the core of the tumor and the invasive margin. This tumor
      block is part of the tumor samples routinely processed by the department of Pathology for
      diagnosis. three tissue sections of 4-microns will be made and deposit on a glass slide to
      perform the immunohistochemistry. Four tissue sections of 4-microns will be made and deposit
      on an Eppendorf safe lock tubes to perform the genetic analyses. Anonymous patient names on
      the slides and tubes will be provided by using a code number and initials of the patient. If
      biopsies for diagnostic purposes are available (i.e. those performed for the patients during
      their initial hospitalization (50% of cases), A tissue sections of each biopsy will be done.
      All slides and tubes will be sent by mail to the HEGP platform for analysis of the immune
      infiltrate and to the Biology department for the genetic analyses. The immunoscore will be
      determine using the following steps: Immunostainings for CD3, CD8, CD45RO on Benchmark XT
      automate; Scanning the immunostainings on Hamamatsu scanner: Quantification of the immune
      populations in specific tumor areas (the core and the invasive margin) with a dedicated image
      analysis module based on the recognition of histological structures adapted to colorectal
      cancer. This module has been developed by our group in connection with the Definiens company.
      A semi-automatic procedure allows the quantification of stained cells in each tumor region.
      Each region is divided into tiles of 800 microns side (500 to 700 tiles analyzed per tumor).
      The density of each marker is calculated using the mean density of the three tiles the most
      infiltrated in each tumor region. A categorization into high density (Hi) or low (Lo) for
      each marker in each tumor region of interest will be performed. This categorization is done
      by comparing the observed density with the optimal cut-off density (min p value approach) we
      previously determined for the DFS and OS in a retrospective study of 250 CRC. Genetic
      analyses: The tubes containing the sections for the genetic analyses of patients with inform
      consent signed for genetic investigation will be transfer to the Department of Biology,
      Hôpital Européen Georges Pompidou. The % of tumor cells of each specimen will be determined
      by a pathologist (based on a H&E evaluation; less than 10%, 10-20%, 20-50%, up to 50% tumor
      cells) and the DNA extraction will be performed. Genetic investigation will comprise the MSI
      status, the presence of a K-Ras and BRAF mutation
observational model: Cohort
time perspective: Prospective
measure: relapse in relation with the immunoscore determined on a tumor section.
time frame: evry 3 month during 3 years and every six months during the fourth year
description: this primary outcome (relapse) will be correlated to the immunoscore determined in the primary tumor (eg. the density of immune cells in tumor regions)
measure: relapse in relation with the immunoscore on biopsies
time frame: evry 3 month during 3 years and every six months during the fourth year
description: this primary outcome (relapse) will be correlated to the immunoscore determined in biopsies performed for diagnosis purpose (eg. the density of immune cells in the biopsies specimens)
measure: relapse in relation with the genetic features of the tumor
time frame: evry 3 month during 3 years and every six months during the fourth year
description: this primary outcome (relapse) will be correlated to the genetic features of the tumor (eg. MSI status, K-Ras and BRAF mutations). The prognostic performance of the genetic and immunologic analyses will be compared.
measure: relapse in relation with the immunological events and psychological status of the patient during the monitoring
time frame: every six months during 4 years
description: a questionnaire will be sent every six months along the monitoring of the patients to obtain informations concerning (i) the emergence of an immune disorder and (ii) the psychological status of the patients. The potential impact of such parameters during the course of the disease on the prediction of the relapse and survival obtained with the immunoscore performed at the time of surgery will be evaluated. The questionnaire should help to identify the clinical parameters to track along the monitoring.
The pathologist of each center will choose a tumor block containing the core of the tumor and
      the invasive margin. Three tissue sections of 4 microns of this tumor block will be cut and
      deposited on glass slides for the immunohistochemical analyses. Four tissue sections of 4
      microns will be cut and deposited in a safe lock eppendorf tube for a DNA extraction. If
      biopsies for diagnostic purpose are available in the laboratory of pathology for the same
      patient, one tissue section of 4 microns of each biopsy will be cut and deposited on a glass
      slide
study pop: 
sampling method: Non-Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: N/A
healthy volunteers: No
last name: Franck Pagès, Professor (MD-PHD)
role: Principal Investigator
affiliation: AP-HP; Paris Descartes University
facility: 
facility: 
facility: 
facility: 
facility: 
facility: 
country: France
citation: Galon J, Costes A, Sanchez-Cabo F, Kirilovsky A, Mlecnik B, Lagorce-Pages C, Tosolini M, Camus M, Berger A, Wind P, Zinzindohoue F, Bruneval P, Cugnenc PH, Trajanoski Z, Fridman WH, Pages F. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science. 2006 Sep 29;313(5795):1960-4. doi: 10.1126/science.1129139.
PMID: 17008531
citation: Pages F, Kirilovsky A, Mlecnik B, Asslaber M, Tosolini M, Bindea G, Lagorce C, Wind P, Marliot F, Bruneval P, Zatloukal K, Trajanoski Z, Berger A, Fridman WH, Galon J. In situ cytotoxic and memory T cells predict outcome in patients with early-stage colorectal cancer. J Clin Oncol. 2009 Dec 10;27(35):5944-51. doi: 10.1200/JCO.2008.19.6147. Epub 2009 Oct 26.
PMID: 19858404
citation: Mlecnik B, Tosolini M, Kirilovsky A, Berger A, Bindea G, Meatchi T, Bruneval P, Trajanoski Z, Fridman WH, Pages F, Galon J. Histopathologic-based prognostic factors of colorectal cancers are associated with the state of the local immune reaction. J Clin Oncol. 2011 Feb 20;29(6):610-8. doi: 10.1200/JCO.2010.30.5425. Epub 2011 Jan 18.
PMID: 21245428
responsible party type: Sponsor
mesh term: Colorectal Neoplasms