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Immunohistochemical Evaluation of Protein P16 Expression in Ovarian Germ Cell Tumors.

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ClinicalTrials.gov Identifier: NCT04283773
Recruitment Status : Completed
First Posted : February 25, 2020
Results First Posted : September 8, 2021
Last Update Posted : September 8, 2021
Sponsor:
Information provided by (Responsible Party):
Omar A Ahmed, Assiut University

Brief Summary:

Ovarian germ cell tumors (OGCTs) constitute 10% of ovarian tumors in Egypt and mainly affect young females. Teratomas are the most common type.Most of teratomas is benign. However, it is liable for malignant transformation. Others are malignant including dysgerminoma, immature teratoma, yolk sac tumor,.etc and accounts 1-1.5% of cancers in young females. The pathogenesis of OGCTs is not clearly understood.

P16 is a member of cyclin-dependent kinase (CDK) inhibitors. It arrests the cell cycle in G1 phase, so it is known as a tumor suppressor protein.P16 immunohistochemical(IHC) expression has been widely investigated in different cancers. Its IHC expression is either absent or overexpressed. Overexpression of p16 is documented in Human Papilloma Virus related endocervical neoplasms and High grade squamous intraepithelial lesions of the vulvovaginal region.Absence of p16 expression is detected in multiple cancers such as Lung cancer, colorectal cancer and lymphoma.

P16 IHC expression in OGCTs is poorly investigated. One study suggests that absent p16 is involved in proliferation of malignant OGCTs via molecular assessment.Another study suggested that decrease P16 is involved in malignant transformation of Mature cystic teratoma to squamous cell carcinoma.However, Previous studies are still limited and recommended further studies to confirm its results.

As the role of altered P16 protein in OGCTs is not widely investigated, we hypothesized that abnormal P16 expression may be involved in its pathogenesis and germ stem cell proliferation.This will give more information about molecular pathways of germ stem cell proliferation to give a hope for CDK inhibitors as novel target therapies in the management of OGCTs.


Condition or disease Intervention/treatment
Ovarian Neoplasms Germ Cell Tumors Immunohistochemistry Combination Product: p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267) Combination Product: Ki67 antibody (DAKO) for malignant cases only

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Study Type : Observational
Actual Enrollment : 62 participants
Observational Model: Case-Control
Time Perspective: Retrospective
Official Title: Immunohistochemical Evaluation of Protein P16 Expression in Ovarian Germ Cell Tumors.
Actual Study Start Date : December 24, 2019
Actual Primary Completion Date : April 1, 2021
Actual Study Completion Date : April 1, 2021


Group/Cohort Intervention/treatment
Malignant ovarian germ cell tumors ( MOGCTs).
22 cases of malignant ovarian germ cell tumors ; include Dysgerminoma , yolk sac tumor and immature teratomas will be treated by anti P16 antibody and Ki67 antibody.
Combination Product: p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267)
Treatment of slides that cut from paraffin embedded blocks related to groups by immunohistochemical method by p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267).

Combination Product: Ki67 antibody (DAKO) for malignant cases only
Treatment of slides that cut from paraffin embedded blocks related to malignant case group by immunohistochemical method by Ki67 antibody.

Mature cystic teratomas.
20 cases of mature teratomas will be treated by anti P16 antibody .
Combination Product: p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267)
Treatment of slides that cut from paraffin embedded blocks related to groups by immunohistochemical method by p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267).

Normally apparent ovaries.
20 cases of normally apparent ovaries will be treated by anti P16 antibody .
Combination Product: p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267)
Treatment of slides that cut from paraffin embedded blocks related to groups by immunohistochemical method by p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267).




Primary Outcome Measures :
  1. P16 Evaluation in Ovarian Germ Cell Tumors [ Time Frame: Antibody exposure overnight, assessed up to 3 days for each run of sections assessed. ]
    For P16 IHC staining, the percentage of P16 positive cells and the location of positive signals (nuclear or cytoplasmic) were visually estimated for neoplastic components of all lesions. German Semi-quantitative scoring system were used to evaluate P16 expression as every tumor will be given a score according to the intensity of the cytoplasmic and nucleic staining (no staining = 0, weak staining = 1, moderate staining = 2, strong staining = 3) And the extent of stained cells (0% = 0, 1-10% = 1, 11-50% =2, 51-80% = 3, 81-100% = 4). The final immunoreactive score will be determined by multiplying the intensity scores with the extent of positivity scores of stained cells, with the minimum score of 0 and a maximum score of 12 ( score 0, 1,2,3,4,6,8,9 and 12).

  2. Measurement of Ki67 Expression in Malignant Ovarian Germ Cell Tumors. [ Time Frame: Antibody exposure 2 hrs , assessed up to 1 day for each run of sections assessed. ]
    For KI67 IHC staining for malignant cases, percentage of nuclear positivity stained cells were assessed, regardless intensity of staining in all sections examined (at least 1000 tumor cells were counted per section for estimation of KI index).Correlation analysis was used to test the association between Ki-67 and other variables (Spearman' Ranked correlation). A p-value < 0.05 was considered significant.


Secondary Outcome Measures :
  1. Correlation Between P16 Cytoplasmic Score and FIGO Staging of MOGCTs. [ Time Frame: After obtaining of results and collecting raw data , within 2 months. ]

    P16 cytoplasmic score is assessed by German quantitative scoring system by multiplying scores of intensity of staining ( 0= No , 1= weak , 2= moderate , 3 = strong ) in scores of intensity of staining ( 0 = less than 10% , 1= 11- 20 % , 2= 21-50 % , 3= 51-80% , 4= more than 80%) to induce final scores range between ( 0, 1,2,4,6,8,9 and 12).

    FIGO staging system for ovarian tumors between Stage I ( limited to ovaries) , II ( with pelvic extension) , III ( with peritoneal extension) and IV ( distant metastasis) , and it is reported from medical records of patients.

    One-way ANOVA was used to examine the Difference in Mean between groups. Post-hoc test with Bonferroni Correction was used for Pairwise comparisons . Data was expressed as mean (SD). A p-value < 0.05 was considered significant.



Biospecimen Retention:   Samples Without DNA

Formalin fixed paraffin embedded blocks of 62 ovarian specimens will be included in this study. These specimens will be divided into 2 groups:

1-42 surgically resected ovarian germ cell tumors specimens: 22 malignant ones( dysgerminoma , immature teratoma and yolk sac tumor) as a case group and equal 20 benign ones (mature cystic teratoma ) as comparison group .

2- 20 normally apparent ovaries that surgically resected with specimens of total abdominal hysterectomy and salpingo-Oopherctomy for non-ovarian causes in perimenopausal women as a control group .



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Ages Eligible for Study:   6 Months to 1 Year   (Child)
Sexes Eligible for Study:   Female
Accepts Healthy Volunteers:   No
Sampling Method:   Probability Sample
Study Population

Sixty-two formalin-fixed, paraffin wax-embedded ovarian specimens .These specimens includes:

(Group A) 22 malignant ovarian germ cell tumors (MOGCTs): 5 dysgerminomas, 8 immature teratomas (four of them Grade II and the other four are Grade III, FIGO grading system) and 9 yolk sac tumors. The patients ranged in age from 12 to 23 years (median age 16.5 years). The initial diagnosis was re-evaluated according to the WHO Classification of Ovarian Tumors.

, (Group B) 20 apparent normal ovarian tissue as a control group that are surgically resected with TAH and salpingo-opherctomy for non-ovarian, non-malignant causes. The patients ranged in age from 42 to 64 years (median 50 years old).

and(Group C) 20 mature cystic teratomas as equal benign comparison group.

Criteria

Inclusion Criteria:

  • Ovarian germ cell tumors :

    1. 20 benign ( mature cystic teratoma ).
    2. 22 malignant ones ( 5 dysgerminoma , 8 immature teratoma and 9 yolk sac tumor).
    3. 20 Normal ovaries .

Exclusion Criteria:

  1. Epithelial ovarian tumors
  2. sex cord -stromal ovarian tumors.
  3. metastatic ovarian lesions.

Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT04283773


Locations
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Egypt
Assiut University
Assiut, Egypt, 71111
Sponsors and Collaborators
Assiut University
Investigators
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Principal Investigator: Omar Ahmed, Master Pathology department- Faculty of medicine - Assiut university
  Study Documents (Full-Text)

Documents provided by Omar A Ahmed, Assiut University:
Study Data/Documents: Clinical Study Report  This link exits the ClinicalTrials.gov site
Treatment outcomes of female germ cell tumors: The Egyptian National Cancer Institute experience. Journal of the Egyptian National Cancer Institute
Clinical Study Report  This link exits the ClinicalTrials.gov site
Incidence and Survival Rates for Female Malignant Germ Cell Tumors
Clinical Study Report  This link exits the ClinicalTrials.gov site
Ovarian Teratomas: Tumor Types and Imaging Characteristics
Clinical Study Report  This link exits the ClinicalTrials.gov site
Early diagnosis of malignant-transformed ovarian mature cystic teratoma: fat-suppressed MRI findings
Clinical Study Report  This link exits the ClinicalTrials.gov site
Significance of β-catenin and pRB pathway components in malignant ovarian germ cell tumours: INK4A promoter CpG island methylation is associated with cell proliferation

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Responsible Party: Omar A Ahmed, Demonstrator at Pathology department - faculty of medicine - Assiut university., Assiut University
ClinicalTrials.gov Identifier: NCT04283773    
Other Study ID Numbers: 17100871
2019-09-05-001-R1 ( Other Grant/Funding Number: Grant office - faculty of medicine - Assiut university )
First Posted: February 25, 2020    Key Record Dates
Results First Posted: September 8, 2021
Last Update Posted: September 8, 2021
Last Verified: August 2021
Individual Participant Data (IPD) Sharing Statement:
Plan to Share IPD: No

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Studies a U.S. FDA-regulated Drug Product: Yes
Studies a U.S. FDA-regulated Device Product: No
Product Manufactured in and Exported from the U.S.: Yes
Keywords provided by Omar A Ahmed, Assiut University:
OGCTs, P16 , Ki67 , IHC
Additional relevant MeSH terms:
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Neoplasms, Germ Cell and Embryonal
Ovarian Neoplasms
Neoplasms by Histologic Type
Neoplasms
Endocrine Gland Neoplasms
Neoplasms by Site
Ovarian Diseases
Adnexal Diseases
Genital Neoplasms, Female
Urogenital Neoplasms
Endocrine System Diseases
Gonadal Disorders
Antibodies
Immunoglobulins
Antibodies, Monoclonal
Immunologic Factors
Physiological Effects of Drugs