Trial Outcomes & Findings for Immunohistochemical Evaluation of Protein P16 Expression in Ovarian Germ Cell Tumors. (NCT NCT04283773)

NCT ID: NCT04283773

Last Updated: 2021-09-08

Results Overview

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).

Recruitment status

COMPLETED

Target enrollment

62 participants

Primary outcome timeframe

Antibody exposure overnight, assessed up to 3 days for each run of sections assessed.

Results posted on

2021-09-08

Participant Flow

1 year .

Participant milestones

Participant milestones
Measure
IHC of P16 Expression in Malignant Ovarian Germ Cell Tumors.
22 cases of malignant ovarian germ cell tumors ; 5cases of dysgerminoma ( one of them is bilateral) , 9 cases of yolk sac tumor and 8 cases of immature teratoma. These cases are treated by anti P16 antibody( p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267) , 1:500 concentration, over night duration and then treated by DAP secondary antibody . Finally assessed by visualized peroxidase method. and also treated by Ki67 antibody.
IHC of P16 Expression in Benign Ovarian Germ Cell Tumors ( Mature Cystic Teratomas).
20 cases of benign ovarian germ cell tumors ( Mature cystic teratomas). These cases are treated by anti P16 antibody( p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267) , 1:500 concentration, over night duration and then treated by DAP secondary antibody.
IHC of P16 Expression in Normal Ovarian Tissues.
20 normal ovarian tissue specimens are used as controls. These cases are treated by anti P16 antibody( p16INK4a Recombinant Rabbit Monoclonal Antibody (RM267) , 1:500 concentration, over night duration and then treated by DAP secondary antibody.
Overall Study
STARTED
22
20
20
Overall Study
COMPLETED
22
20
20
Overall Study
NOT COMPLETED
0
0
0

Reasons for withdrawal

Withdrawal data not reported

Baseline Characteristics

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

Baseline characteristics by cohort

Baseline characteristics by cohort
Measure
Malignant Ovarian Germ Cell Tumors ( MOGCTs).
n=22 Participants
22 cases of malignant ovarian germ cell tumors ; include Dysgerminoma , yolk sac tumor and immature teratomas will be treated by anti P16 antibody . 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).
Mature Cystic Teratomas.
n=20 Participants
20 cases of mature teratomas will be treated by anti P16 antibody .
Normally Apparent Ovaries.
n=20 Participants
20cases of normally apparent ovaries will be treated by anti P16 antibody .
Total
n=62 Participants
Total of all reporting groups
Age, Continuous
16.5 years
n=39 Participants
31.5 years
n=41 Participants
50 years
n=35 Participants
30 years
n=31 Participants
Sex: Female, Male
Female
22 Participants
n=39 Participants
20 Participants
n=41 Participants
20 Participants
n=35 Participants
62 Participants
n=31 Participants
Sex: Female, Male
Male
0 Participants
n=39 Participants
0 Participants
n=41 Participants
0 Participants
n=35 Participants
0 Participants
n=31 Participants
Ethnicity (NIH/OMB)
Hispanic or Latino
0 Participants
n=39 Participants
0 Participants
n=41 Participants
0 Participants
n=35 Participants
0 Participants
n=31 Participants
Ethnicity (NIH/OMB)
Not Hispanic or Latino
22 Participants
n=39 Participants
20 Participants
n=41 Participants
20 Participants
n=35 Participants
62 Participants
n=31 Participants
Ethnicity (NIH/OMB)
Unknown or Not Reported
0 Participants
n=39 Participants
0 Participants
n=41 Participants
0 Participants
n=35 Participants
0 Participants
n=31 Participants
Region of Enrollment
Egypt
22 participants
n=39 Participants
20 participants
n=41 Participants
20 participants
n=35 Participants
62 participants
n=31 Participants
P16 cytoplasmic expression
7 units on a scale
n=39 Participants
4 units on a scale
n=41 Participants
0 units on a scale
n=35 Participants
6 units on a scale
n=31 Participants
P16 nuclear expression
6 units on a scale
n=39 Participants
4 units on a scale
n=41 Participants
0 units on a scale
n=35 Participants
4 units on a scale
n=31 Participants

PRIMARY outcome

Timeframe: 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).

Outcome measures

Outcome measures
Measure
Malignant Ovarian Germ Cell Tumors ( MOGCTs).
n=22 Participants
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. 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Mature Cystic Teratomas.
n=20 Participants
20 cases of mature teratomas will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Normally Apparent Ovaries.
n=20 Participants
20 cases of normally apparent ovaries will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
P16 Evaluation in Ovarian Germ Cell Tumors
P16 cytoplasmic expression
7 units on a scale
Interval 0.0 to 8.0
4 units on a scale
Interval 0.0 to 6.0
0 units on a scale
Interval 0.0 to 4.0
P16 Evaluation in Ovarian Germ Cell Tumors
P16 nuclear expression
6 units on a scale
Interval 0.0 to 8.0
4 units on a scale
Interval 0.0 to 5.0
0 units on a scale
Interval 0.0 to 3.0

PRIMARY outcome

Timeframe: Antibody exposure 2 hrs , assessed up to 1 day for each run of sections assessed.

Population: Measurement of Ki67 expression in malignant ovarian germ cell tumors only. That explains why analysis obtained on Arm/Group 1 ( MOGCTs) only.

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.

Outcome measures

Outcome measures
Measure
Malignant Ovarian Germ Cell Tumors ( MOGCTs).
n=22 Participants
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. 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Mature Cystic Teratomas.
20 cases of mature teratomas will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Normally Apparent Ovaries.
20 cases of normally apparent ovaries will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Measurement of Ki67 Expression in Malignant Ovarian Germ Cell Tumors.
15 percentage of Ki67 positive nuclear cell
Interval 0.0 to 100.0

SECONDARY outcome

Timeframe: After obtaining of results and collecting raw data , within 2 months.

Population: 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 for (malignant ovarian tumors only) is reported from medical records of patients . That explains why this measurement Arm/group 1( MOGCTs)only.

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.

Outcome measures

Outcome measures
Measure
Malignant Ovarian Germ Cell Tumors ( MOGCTs).
n=22 Participants
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. 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Mature Cystic Teratomas.
20 cases of mature teratomas will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Normally Apparent Ovaries.
20 cases of normally apparent ovaries will be treated by anti P16 antibody . 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).Additional step for malignant cases , sections were treated by Ki67 antibody.
Correlation Between P16 Cytoplasmic Score and FIGO Staging of MOGCTs.
11 score on a scale
Interval 9.5 to 13.5

Adverse Events

Immunohistochemical Expression of P16 Expression in Malignant Ovarian Germ Cell Tumors.

Serious events: 0 serious events
Other events: 0 other events
Deaths: 0 deaths

Immunohistochemical Expression of P16 Expression in Benign Ovarian Germ Cell (MCTs)

Serious events: 0 serious events
Other events: 0 other events
Deaths: 0 deaths

Immunohistochemical Expression of P16 Expression in Normal Ovarian Tissues.

Serious events: 0 serious events
Other events: 0 other events
Deaths: 0 deaths

Serious adverse events

Adverse event data not reported

Other adverse events

Adverse event data not reported

Additional Information

Demonstrator at pathology department

Assiut Univercity

Phone: 00201033986747

Results disclosure agreements

  • Principal investigator is a sponsor employee
  • Publication restrictions are in place