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Marker genes for oocyte competence

A technology of oocytes and cumulus cells, which is applied in the field of end-point standards of oocytes, and can solve problems such as undiscovered genes

Pending Publication Date: 2015-12-16
VRIJE UNIV BRUSSEL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current literature, many genes that are common across different studies were not identified

Method used

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  • Marker genes for oocyte competence
  • Marker genes for oocyte competence
  • Marker genes for oocyte competence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-Q

[0166] Embodiment 1-QPCR analysis

[0167] Materials and Methods

[0168] Patient group

[0169] This study was approved by the UZBrussel Ethics Committee and written consent was obtained from the patients. Forty-seven ICSI patients were selected based on the embryo transfer policy (single embryo transfer), and an ovulation induction protocol was formulated: GnRH antagonist (Gonal-f, Merck-Serono, Geneva, Switzerland; n=4 or Puregon, MSD, Oss, Netherlands; n=43). Causes of infertility were: male factor only (n=19), female factor only (ovulation disorder n=3, and tubal infertility n=2), combined male and female factor (OAT and endometriosis n =3) and idiopathic (n=20). Twenty patients had single embryo transfers at culture day 3, of which 10 became pregnant. Twenty-seven patients had transplants at day 5, of which 9 became pregnant.

[0170] Collection of human cumulus cells and culture of embryos

[0171] Vaginal ultrasound is used to monitor follicular development. Th...

Embodiment 2

[0238] Example 2 - Exon level analysis

[0239] Three microarray experiments were performed and the resulting data resulted in 4 microarray analyses. Designated below as Microarray Analysis 1, 2, 3 or 4, respectively (see Table 14 below). Array experiments 1 and 2 were used for data mining (to find genes or exons of genes that were predictive for oocyte competence), and array experiment 3 was used to filter frequently occurring predictive genes (to find genes that could be applied more generally or exons, as they were predictive in different fertility centers and after slightly different treatments). The most recent analysis is based on a microarray experiment 2 that includes intra-patient and intra-cycle (oocytes from the same stimulated cohort) analysis for live births.

[0240] Material:

[0241] For the three microarray experiments, cumulus cells were collected as previously described (Wathlet et al. 2011). RNA was extracted using the Qiagen Rneasymicro kit as descri...

Embodiment 3

[0275] Example 3 - Retrospective Study

[0276] A retrospective study was performed with samples collected from 107 patients in two clinical settings. QPCR for 7 oocyte quality markers (HSPH1 exon 2, HSPH1 exon 6, CAMK1D exon 1, CAMK1D exon 9, EFNB2, NCOA7, Exon execution specific to SASH1) and 2 endogenous controls (UBC and B2M) (Table 9).

[0277] Table 9: Samples available for proof-of-concept studies. B1 / 2 / 3 = Batch 1 / 2 / 3, Site 1+2 = European and US Fertility Centers, Patient Pretreatment = GnRH antagonist in combination with rFSH or HP-hMG, and Pregnancy Outcome = At ​​least 2 pregnancies post-transplant weeks, non-pregnant = no pregnancy after transfer, delayed = embryos developed slowly and not considered for transfer

[0278]

[0279] Cumulus cell collection:

[0280] Cumulus cell samples were collected at one European and one US fertility center (Table 9) and were derived from patients who had obtained one embryo / transfer (SET = single embryo transfer) and de...

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Abstract

Cumulus cell (CC) gene expression is explored as an additional method to morphological scoring to choose the embryo with the highest chance to pregnancy. The present invention relates to a novel method of identifying biomarker genes for evaluating the competence of a mammalian oocyte in giving rise to a viable pregnancy after fertilization, based on the use of live birth and embryonic development as endpoint criteria for the oocytes to be used in an exon level analysis of potential biomarker genes. The invention further provides CC-expressed biomarker genes thus identified, as well as prognostic models based on the biomarker genes identified using the methods of the present invention.

Description

field of invention [0001] Cumulus cell (CC) gene expression is being explored as an additional method of morphological scoring to select embryos with the highest chance of pregnancy. The present invention relates to a novel method for the identification of biomarker genes for assessing the ability of mammalian oocytes to produce a viable pregnancy after fertilization, based on the use of live birth and embryonic development as an alternative to potential biomarker genes. Endpoint criteria for oocytes to be used in exome-level analysis. The present invention further provides CC expressed biomarker genes thus identified, as well as prognostic models based on the biomarker genes identified using the methods of the present invention. Background of the invention [0002] Single embryo transfer (SET) is the preferred management to limit multiple pregnancies after ART. In order not to jeopardize the take-home baby rate, the selection of embryos for transfer in the first cycle bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q2600/158C12Q1/6881C12N5/0609C12Q2600/124
Inventor J·史密兹S·沃斯勒特T·阿德里恩森斯
Owner VRIJE UNIV BRUSSEL
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