Ignite Creation Date:
2024-05-06 @ 10:51 AM
Last Modification Date:
2024-10-26 @ 12:36 PM
Study NCT ID:
NCT03361839
Status:
UNKNOWN
Last Update Posted:
2017-12-05
First Post:
2017-11-18
Brief Title:
Endometrial Receptivity in Patients With Recurrent Implantation Failure as a Preparatory Step for PET
Sponsor:
Cairo University
Organization:
Cairo University
Study Overview
Official Title:
Clinical Trail for Study Endometrial Receptivity in RIF
Status:
UNKNOWN
Status Verified Date:
2017-12
Last Known Status:
NOT_YET_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:
clinical trial to improve pregnancy outcome in patient with RLF study done on 2 phases phase 1 to study variations of endometrial receptivity genes in this group of patient phase 2 repeat cycle or transfer frozen embryo in proper time and after giving proper treatment
Detailed Description:
Study design Nonrandomized clinical trial
Inclusion criteria women with history of recurrent implantation failure with History of transfer of at least 4 good quality embryos in at least 3 fresh or frozen cycles
Women age less than 40 years
BMI 25 - 33
Non diabetic Arms of study 2
1 Women for frozen embryo transfer or New ICSI cycle fulfilling previous criteria study arm
2 Control group of fertile patient fertile patients receiving local mechanical contraception as a reference group To help in identification of cut off values of all related genes
Sample size 60 Intervention
Phase 1
-- --
1 Recent labs day 2 serum FSHLH TSH PROLACTIN serum E 2within 6 ms
2 White tablets of cycloprogenova 4 mg 12 hs twice day
3 Add progesterone on day 9 600 mg per day
4 On day 5 of start progesterone do endometrial biopsy using pipplle endometrial suction curette
5 Indirect immunofluorescence using QRT-PCR of the endometrial tissue for 11 genes necessary for implantation and maintenance of pregnancy Classify EB as pre-receptive receptive and post receptive and plan next day of transfer
6 Genes to be studied the most accessible genes will be studied
-- --
1 Homeobox gene HOXA-10 upregulated
2 LIF upregulated
3 Alpha VB3 integren and its ligand osteopontin are positively detected
4 ECM upregulated
5 Paracrine stromal factors ve EGF epidermal growth factor heparin binding EGF
VE 17 B estradiol E2 factors 6- L selectin upregulated 7- E-cadherin downregulated 8- Intercellular cell adhesion molecules highly expressed 9- Mucin 1 down regulated at implantation 10- IL-6 upregulated 11- IL-1 and IL- 1 R upregulated 12- Prostaglandin transporter PGT reduced in mid-late secretory Phase 2 7- In the study arm Next cycle start active ttt cycle of frozen ET white tablet of Cycloprogenova twice daily then at day 9 do transvaginal US when endometrial thickness is more than 9 mm start progesterone till day 5 for transfer of frozen embryos or new ICSI cycle according to standard protocol long or antagonist protocols according to every patient data then select one of the suggested treatment
1 Intrauterine injection of human chorionic gonadotropins before transfer
2 Granulocyte colony stimulating factor given intrauterine 300 mcg ml on day of OP or progesterone administration of FET
3 Recombinant LIF
4 Or combination of 1 and 2 Common measures to be done in any situation
-- --
1 Low dose aspirin 75 mg daily
2 Steroids 5 mg hostacortin daily
3 Endometrial injury cycle before active cycle treatment
8- Technique of QRT-PCR of endometrial receptivity genes RNA Extraction Cells of all studied groups will be lysed and total RNA was isolated with RNAeasy Mini Kit Qiagen and further analyzed for quantity and quality with Beckman dual spectrophotometer USA
Real Time PCR qRT-PCR For quantitative expression of HOXA-10 LIF Alpha VB3 integren and its ligand osteopontin ECM EGF E2 FACTOR L-selectin E-cadherin ICAM Mucin1 IL-6 IL1 IL-1R and PGT the following procedure will bes assessed 10 ng of the total RNA from each sample will be used for cDNA synthesis by reverse transcription using High capacity cDNA Reverse Transcriptase kit Applied Biosystem USA The cDNA will be subsequently amplified with the Syber Green I PCR Master Kit Fermentas in a 48-well plate using the Step One instrument Applied Biosystem USA as follows 10 minutes at 95 ºC for enzyme activation followed by 40 cycles of 15 seconds at 95ºC 20 seconds at 55 ºC and 30 second at 72 ºC for the amplification step Changes in the expression of each target gene will be normalized relative to the mean critical threshold CT values of β-actin as housekeeping gene by the ΔΔCt method We will use 1 μM of both primers specific for each target gene DNA sequencing will be assessed for all studied genes
Inclusion criteria women with history of recurrent implantation failure with History of transfer of at least 4 good quality embryos in at least 3 fresh or frozen cycles
Women age less than 40 years
BMI 25 - 33
Non diabetic Arms of study 2
1 Women for frozen embryo transfer or New ICSI cycle fulfilling previous criteria study arm
2 Control group of fertile patient fertile patients receiving local mechanical contraception as a reference group To help in identification of cut off values of all related genes
Sample size 60 Intervention
Phase 1
-- --
1 Recent labs day 2 serum FSHLH TSH PROLACTIN serum E 2within 6 ms
2 White tablets of cycloprogenova 4 mg 12 hs twice day
3 Add progesterone on day 9 600 mg per day
4 On day 5 of start progesterone do endometrial biopsy using pipplle endometrial suction curette
5 Indirect immunofluorescence using QRT-PCR of the endometrial tissue for 11 genes necessary for implantation and maintenance of pregnancy Classify EB as pre-receptive receptive and post receptive and plan next day of transfer
6 Genes to be studied the most accessible genes will be studied
-- --
1 Homeobox gene HOXA-10 upregulated
2 LIF upregulated
3 Alpha VB3 integren and its ligand osteopontin are positively detected
4 ECM upregulated
5 Paracrine stromal factors ve EGF epidermal growth factor heparin binding EGF
VE 17 B estradiol E2 factors 6- L selectin upregulated 7- E-cadherin downregulated 8- Intercellular cell adhesion molecules highly expressed 9- Mucin 1 down regulated at implantation 10- IL-6 upregulated 11- IL-1 and IL- 1 R upregulated 12- Prostaglandin transporter PGT reduced in mid-late secretory Phase 2 7- In the study arm Next cycle start active ttt cycle of frozen ET white tablet of Cycloprogenova twice daily then at day 9 do transvaginal US when endometrial thickness is more than 9 mm start progesterone till day 5 for transfer of frozen embryos or new ICSI cycle according to standard protocol long or antagonist protocols according to every patient data then select one of the suggested treatment
1 Intrauterine injection of human chorionic gonadotropins before transfer
2 Granulocyte colony stimulating factor given intrauterine 300 mcg ml on day of OP or progesterone administration of FET
3 Recombinant LIF
4 Or combination of 1 and 2 Common measures to be done in any situation
-- --
1 Low dose aspirin 75 mg daily
2 Steroids 5 mg hostacortin daily
3 Endometrial injury cycle before active cycle treatment
8- Technique of QRT-PCR of endometrial receptivity genes RNA Extraction Cells of all studied groups will be lysed and total RNA was isolated with RNAeasy Mini Kit Qiagen and further analyzed for quantity and quality with Beckman dual spectrophotometer USA
Real Time PCR qRT-PCR For quantitative expression of HOXA-10 LIF Alpha VB3 integren and its ligand osteopontin ECM EGF E2 FACTOR L-selectin E-cadherin ICAM Mucin1 IL-6 IL1 IL-1R and PGT the following procedure will bes assessed 10 ng of the total RNA from each sample will be used for cDNA synthesis by reverse transcription using High capacity cDNA Reverse Transcriptase kit Applied Biosystem USA The cDNA will be subsequently amplified with the Syber Green I PCR Master Kit Fermentas in a 48-well plate using the Step One instrument Applied Biosystem USA as follows 10 minutes at 95 ºC for enzyme activation followed by 40 cycles of 15 seconds at 95ºC 20 seconds at 55 ºC and 30 second at 72 ºC for the amplification step Changes in the expression of each target gene will be normalized relative to the mean critical threshold CT values of β-actin as housekeeping gene by the ΔΔCt method We will use 1 μM of both primers specific for each target gene DNA sequencing will be assessed for all studied genes
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None