<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/NCT03002727</url>
  </required_header>
  <id_info>
    <org_study_id>2016/06/17-002</org_study_id>
    <nct_id>NCT03002727</nct_id>
  </id_info>
  <brief_title>Role of CD133 &amp; Microsatellite Status in Evaluation of Rectosigmoid Cancer Young Adults Received Neoadjuvant Treatment</brief_title>
  <official_title>Role of CD133 and Microsatellite Status in Evaluation of Young Adults With Rectosigmoid Cancer Received Neoadjuvant Treatment</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Assiut University</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>Assiut University</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Microsatellite instability is more common in colorectal cancer ( CRC) young patient which is&#xD;
      associated with good prognosis and is considered as a predictor for good response to&#xD;
      preoperative chemoradiotherapy. Counting of ( cluster of differentiation) CD 133 +ve cells&#xD;
      ,as a marker for enrichment with colorectal cancer stem cells ,is considered as a prognostic&#xD;
      marker for poor survival and predictor for radio-resistance. Correlation between&#xD;
      microsatellite status ( MS) and CD133 count has not yet studied especially in young patients&#xD;
      with rectosigmoid cancer. So the investigators hypothesize that there is correlation between&#xD;
      microsatellite status, CD133+ve cells count , occurrence of CRC in young patients and&#xD;
      resistance to standard treatment regimen. Improvement of response to treatment and choice of&#xD;
      the best regime to avoid non beneficial treatment modality are the goal of this study.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      The age incidence of colorectal cancer is above fifty years old world wide .But, when&#xD;
      comparing the incidence and clinicopathological features of colorectal cancer in western&#xD;
      countries and countries with constrained resources as Egypt , there is significant lower&#xD;
      median age of incidence. In Egypt reports showed that CRC was detected in 11-15% of patients&#xD;
      who underwent colonoscopy and diagnosed in 29-31% of patients aged 40 years or younger .&#xD;
      Similar data come from other highly populated resource-constrained countries in Asia . This&#xD;
      early onset rectal cancer is mainly poorly differentiated, advanced at prognosis, sporadic&#xD;
      with no familial predisposition.Young patients with microsatellite high (MSI-h ) proximal&#xD;
      cancer colon are with good prognosis but there is lacking data about the prognostic and&#xD;
      predictive role of these genes in young patients with irradiated rectal cancer. As those&#xD;
      young aged patients show worse outcome in term of progression free survival and higher rates&#xD;
      of metastatic events , we hypothesize that colorectal cancer stem cells (CR-CSC) may have a&#xD;
      role in this dismal outcome.CD133 is one of the best-characterized markers of CR-CSCs, many&#xD;
      studies have demonstrated that CD133 expression was correlated with survival, recurrence,&#xD;
      metastasis and chemotherapy resistance in colorectal cancer.&#xD;
&#xD;
      So the study is trying to establish a correlation between the MS status and enrichment with&#xD;
      CR-CSC and if this proposed relationship may have implication of disease outcome and&#xD;
      treatment results.&#xD;
&#xD;
      Patients and methods:&#xD;
&#xD;
      The investigators will enroll about 30 patients and will examine the pre- treatment&#xD;
      colonoscopic biopsy for&#xD;
&#xD;
        -  Type and grade of carcinoma under light microscopy after staining with haematoxylin and&#xD;
           eosin.&#xD;
&#xD;
        -  Immunocytochemistry performed on 4 µ-thick sections from paraffin blocks using the&#xD;
           streptavidin-biotin-peroxidase technique. The sections will be routinely deparaffinized,&#xD;
           rehydrated through graded alcohols to distilled water. Deparaffinized sections will be&#xD;
           treated with 0.3% hydrogen peroxide for 10 min. Suitable antigen retrieval will be&#xD;
           carried out. Blocking serum will be applied for 10 min. The sections will be incubated&#xD;
           with antibody raised against&#xD;
&#xD;
             1. MMR proteins : MLH1, MSH2, or MSH6,PMS-2 .&#xD;
&#xD;
             2. CD133. Then the investigators will collect the blocks from the same patients after&#xD;
                receiving neoadjuvant chemoradiotherapy and underwent surgery and examine&#xD;
&#xD;
             1. Under light microscope after staining for :Type and grade of carcinoma&#xD;
                Angiovascular invasion Degree of immune response Pattern and depth of invasion&#xD;
                Lymph node status.&#xD;
&#xD;
             2. CD 133 positive cell count with the same previously reported technique . The&#xD;
                resulting immune complex will be detected by a universal staining kit following the&#xD;
                instructions attached with the kit. Slides will be washed several times in&#xD;
                phosphate buffer saline and will be placed in it for 5 minutes between each step&#xD;
                except after peroxidase reagent the washing will be by distilled water. Negative&#xD;
                and positive controls will be included in each staining series. Immunohistochemical&#xD;
                evaluation will be done according to Ren F et al and Zhang X et al . Digital images&#xD;
                will be obtained with a digital camera system&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Unknown status</overall_status>
  <last_known_status>Recruiting</last_known_status>
  <start_date>December 2016</start_date>
  <completion_date type="Anticipated">January 2019</completion_date>
  <primary_completion_date type="Anticipated">December 2018</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>Differential expression of CD133 based on microsatellite status and correlation with clinicopathological features.</measure>
    <time_frame>3 month</time_frame>
  </primary_outcome>
  <secondary_outcome>
    <measure>Assess the microsatellite status of young adult with rectosigmoid cancer</measure>
    <time_frame>3 months</time_frame>
    <description>Define the microsatellite status of this group of patient as it is underrepresented in the most of studies</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>The effect of radiotherapy on the count of CD 133 +ve cells</measure>
    <time_frame>3 months</time_frame>
    <description>Examine if radiotherapy increases the stemness of the tumour</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Define microsatellite status and enrichment with CD + ve 133 as colorectal cancer stem cells as a prognostic marker for this group of patients</measure>
    <time_frame>6 months</time_frame>
  </secondary_outcome>
  <secondary_outcome>
    <measure>If radiotherapy increases the count of CD +ve 133 cells , does that increase significantly affects the progression free survival</measure>
    <time_frame>6 month</time_frame>
  </secondary_outcome>
  <enrollment type="Anticipated">30</enrollment>
  <condition>Colorectal Cancer Genetics</condition>
  <condition>Young Adult</condition>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      Paraffin blocks of colonoscopic biobsy before neoadjuvant concurrent chemoradiotherapy and&#xD;
      the blocks of surgery at the end of treatment regemin.&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Patients aged less than fourty years old with rectosigmoid cancer diagnosed with&#xD;
        colonscopic biobsy and received concurrent chemoradiotherapy then went for surgery&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Non-Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Patient less than forty years old with rectosigmoid cancer.&#xD;
&#xD;
          -  Sporadic colorectal cancer patients; no family history of first degree relatives .&#xD;
&#xD;
          -  Diagnosed with colonoscopic biopsy.&#xD;
&#xD;
          -  Patient received neoadjuvant concurrent chemoradiotherapy.&#xD;
&#xD;
          -  Radical surgery was done after neoadjuvant treatment .&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Patient less than 18 and more than forty.&#xD;
&#xD;
          -  Patient who underwent radical surgery from the start for early disease.&#xD;
&#xD;
          -  Patient with metastatic colorectal cancer.&#xD;
&#xD;
          -  patients with positive family history.&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>40 Years</maximum_age>
    <healthy_volunteers>No</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Ola N Abdl fattah, Ass.Prof</last_name>
    <role>Study Director</role>
    <affiliation>Assiut University</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Dalia A El sers, Ass.prof</last_name>
    <role>Study Director</role>
    <affiliation>Assiut University</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Mahmoud R Shehata, Lecturer</last_name>
    <role>Study Director</role>
    <affiliation>Assiut University</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Samia A Ali, Prof</last_name>
    <role>Study Chair</role>
    <affiliation>Assiut University</affiliation>
  </overall_official>
  <overall_contact>
    <last_name>Ereny S Poles, MsC</last_name>
    <phone>01000470421</phone>
    <phone_ext>+2</phone_ext>
    <email>Rerejesus_dna@yahoo.com</email>
  </overall_contact>
  <location>
    <facility>
      <name>Ereny Samwel Poles</name>
      <address>
        <city>Assiut</city>
        <country>Egypt</country>
      </address>
    </facility>
    <status>Recruiting</status>
    <contact>
      <last_name>Ereny S Poles, MsC</last_name>
      <phone>01000470421</phone>
      <phone_ext>+2</phone_ext>
      <email>rerejesus_dna@yahoo.com</email>
    </contact>
    <investigator>
      <last_name>Ola N Abdl fatah, Ass.prof</last_name>
      <role>Sub-Investigator</role>
    </investigator>
  </location>
  <location_countries>
    <country>Egypt</country>
  </location_countries>
  <reference>
    <citation>Chiang JM, Chen MC, Changchien CR, Chen JS, Tang R, Wang JY, Yeh CY, Fan CW, Tsai WS. Favorable influence of age on tumor characteristics of sporadic colorectal adenocarcinoma: patients 30 years of age or younger may be a distinct patient group. Dis Colon Rectum. 2003 Jul;46(7):904-10. doi: 10.1007/s10350-004-6683-1.</citation>
    <PMID>12847364</PMID>
  </reference>
  <reference>
    <citation>El-Hennawy MM, Moussa ME, El-Saeidy MK, Shawky AM, Bessa SS, Badour NM. Rectal carcinoma in Egyptian patients less than 40 years of age. Int Surg. 2003 Jul-Sep;88(3):137-44.</citation>
    <PMID>14584768</PMID>
  </reference>
  <reference>
    <citation>Park CH, Kim HC, Cho YB, Yun SH, Lee WY, Park YS, Choi DH, Chun HK. Predicting tumor response after preoperative chemoradiation using clinical parameters in rectal cancer. World J Gastroenterol. 2011 Dec 28;17(48):5310-6. doi: 10.3748/wjg.v17.i48.5310.</citation>
    <PMID>22219601</PMID>
  </reference>
  <results_reference>
    <citation>Huh JW, Kim HC, Kim SH, Park YA, Cho YB, Yun SH, Lee WY, Park HC, Choi DH, Park JO, Park YS, Chun HK. Mismatch Repair Gene Expression as a Predictor of Tumor Responses in Patients With Rectal Cancer Treated With Preoperative Chemoradiation. Medicine (Baltimore). 2016 Jan;95(3):e2582. doi: 10.1097/MD.0000000000002582.</citation>
    <PMID>26817916</PMID>
  </results_reference>
  <verification_date>May 2018</verification_date>
  <study_first_submitted>December 21, 2016</study_first_submitted>
  <study_first_submitted_qc>December 23, 2016</study_first_submitted_qc>
  <study_first_posted type="Estimate">December 26, 2016</study_first_posted>
  <last_update_submitted>May 5, 2018</last_update_submitted>
  <last_update_submitted_qc>May 5, 2018</last_update_submitted_qc>
  <last_update_posted type="Actual">May 8, 2018</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>Assiut University</investigator_affiliation>
    <investigator_full_name>Ereny Samwel</investigator_full_name>
    <investigator_title>Assistant lecturer</investigator_title>
  </responsible_party>
  <keyword>Colorectal cancer</keyword>
  <keyword>Microsatellite</keyword>
  <keyword>Colorectal cancer stem cells</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Colorectal Neoplasms</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>No</sharing_ipd>
  </patient_data>
  <!-- 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/NCT03002727
org study id: 2016/06/17-002
nct id: NCT03002727
lead sponsor: 
has dmc: No
Microsatellite instability is more common in colorectal cancer ( CRC) young patient which is
      associated with good prognosis and is considered as a predictor for good response to
      preoperative chemoradiotherapy. Counting of ( cluster of differentiation) CD 133 +ve cells
      ,as a marker for enrichment with colorectal cancer stem cells ,is considered as a prognostic
      marker for poor survival and predictor for radio-resistance. Correlation between
      microsatellite status ( MS) and CD133 count has not yet studied especially in young patients
      with rectosigmoid cancer. So the investigators hypothesize that there is correlation between
      microsatellite status, CD133+ve cells count , occurrence of CRC in young patients and
      resistance to standard treatment regimen. Improvement of response to treatment and choice of
      the best regime to avoid non beneficial treatment modality are the goal of this study.
The age incidence of colorectal cancer is above fifty years old world wide .But, when
      comparing the incidence and clinicopathological features of colorectal cancer in western
      countries and countries with constrained resources as Egypt , there is significant lower
      median age of incidence. In Egypt reports showed that CRC was detected in 11-15% of patients
      who underwent colonoscopy and diagnosed in 29-31% of patients aged 40 years or younger .
      Similar data come from other highly populated resource-constrained countries in Asia . This
      early onset rectal cancer is mainly poorly differentiated, advanced at prognosis, sporadic
      with no familial predisposition.Young patients with microsatellite high (MSI-h ) proximal
      cancer colon are with good prognosis but there is lacking data about the prognostic and
      predictive role of these genes in young patients with irradiated rectal cancer. As those
      young aged patients show worse outcome in term of progression free survival and higher rates
      of metastatic events , we hypothesize that colorectal cancer stem cells (CR-CSC) may have a
      role in this dismal outcome.CD133 is one of the best-characterized markers of CR-CSCs, many
      studies have demonstrated that CD133 expression was correlated with survival, recurrence,
      metastasis and chemotherapy resistance in colorectal cancer.

      So the study is trying to establish a correlation between the MS status and enrichment with
      CR-CSC and if this proposed relationship may have implication of disease outcome and
      treatment results.

      Patients and methods:

      The investigators will enroll about 30 patients and will examine the pre- treatment
      colonoscopic biopsy for

        -  Type and grade of carcinoma under light microscopy after staining with haematoxylin and
           eosin.

        -  Immunocytochemistry performed on 4 µ-thick sections from paraffin blocks using the
           streptavidin-biotin-peroxidase technique. The sections will be routinely deparaffinized,
           rehydrated through graded alcohols to distilled water. Deparaffinized sections will be
           treated with 0.3% hydrogen peroxide for 10 min. Suitable antigen retrieval will be
           carried out. Blocking serum will be applied for 10 min. The sections will be incubated
           with antibody raised against

             1. MMR proteins : MLH1, MSH2, or MSH6,PMS-2 .

             2. CD133. Then the investigators will collect the blocks from the same patients after
                receiving neoadjuvant chemoradiotherapy and underwent surgery and examine

             1. Under light microscope after staining for :Type and grade of carcinoma
                Angiovascular invasion Degree of immune response Pattern and depth of invasion
                Lymph node status.

             2. CD 133 positive cell count with the same previously reported technique . The
                resulting immune complex will be detected by a universal staining kit following the
                instructions attached with the kit. Slides will be washed several times in
                phosphate buffer saline and will be placed in it for 5 minutes between each step
                except after peroxidase reagent the washing will be by distilled water. Negative
                and positive controls will be included in each staining series. Immunohistochemical
                evaluation will be done according to Ren F et al and Zhang X et al . Digital images
                will be obtained with a digital camera system
observational model: Cohort
time perspective: Prospective
measure: Differential expression of CD133 based on microsatellite status and correlation with clinicopathological features.
time frame: 3 month
measure: Assess the microsatellite status of young adult with rectosigmoid cancer
time frame: 3 months
description: Define the microsatellite status of this group of patient as it is underrepresented in the most of studies
measure: The effect of radiotherapy on the count of CD 133 +ve cells
time frame: 3 months
description: Examine if radiotherapy increases the stemness of the tumour
measure: Define microsatellite status and enrichment with CD + ve 133 as colorectal cancer stem cells as a prognostic marker for this group of patients
time frame: 6 months
measure: If radiotherapy increases the count of CD +ve 133 cells , does that increase significantly affects the progression free survival
time frame: 6 month
Paraffin blocks of colonoscopic biobsy before neoadjuvant concurrent chemoradiotherapy and
      the blocks of surgery at the end of treatment regemin.
study pop: 
sampling method: Non-Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: 40 Years
healthy volunteers: No
last name: Ola N Abdl fattah, Ass.Prof
role: Study Director
affiliation: Assiut University
last name: Dalia A El sers, Ass.prof
role: Study Director
affiliation: Assiut University
last name: Mahmoud R Shehata, Lecturer
role: Study Director
affiliation: Assiut University
last name: Samia A Ali, Prof
role: Study Chair
affiliation: Assiut University
last name: Ereny S Poles, MsC
phone: 01000470421
phone ext: +2
email: Rerejesus_dna@yahoo.com
facility: 
status: Recruiting
contact: 
investigator: 
country: Egypt
citation: Chiang JM, Chen MC, Changchien CR, Chen JS, Tang R, Wang JY, Yeh CY, Fan CW, Tsai WS. Favorable influence of age on tumor characteristics of sporadic colorectal adenocarcinoma: patients 30 years of age or younger may be a distinct patient group. Dis Colon Rectum. 2003 Jul;46(7):904-10. doi: 10.1007/s10350-004-6683-1.
PMID: 12847364
citation: El-Hennawy MM, Moussa ME, El-Saeidy MK, Shawky AM, Bessa SS, Badour NM. Rectal carcinoma in Egyptian patients less than 40 years of age. Int Surg. 2003 Jul-Sep;88(3):137-44.
PMID: 14584768
citation: Park CH, Kim HC, Cho YB, Yun SH, Lee WY, Park YS, Choi DH, Chun HK. Predicting tumor response after preoperative chemoradiation using clinical parameters in rectal cancer. World J Gastroenterol. 2011 Dec 28;17(48):5310-6. doi: 10.3748/wjg.v17.i48.5310.
PMID: 22219601
citation: Huh JW, Kim HC, Kim SH, Park YA, Cho YB, Yun SH, Lee WY, Park HC, Choi DH, Park JO, Park YS, Chun HK. Mismatch Repair Gene Expression as a Predictor of Tumor Responses in Patients With Rectal Cancer Treated With Preoperative Chemoradiation. Medicine (Baltimore). 2016 Jan;95(3):e2582. doi: 10.1097/MD.0000000000002582.
PMID: 26817916
responsible party type: Principal Investigator
investigator affiliation: Assiut University
investigator full name: Ereny Samwel
investigator title: Assistant lecturer
mesh term: Colorectal Neoplasms
sharing ipd: No