Methods for selecting competent oocytes and competent embryos with high potential for pregnancy outcome

Patent metadata

Field	Value
Publication number	US-10059994-B2
Application number	US-201615356745-A
Country	US
Kind code	B2
Filing date	Nov 21, 2016
Priority date	Nov 24, 2010
Publication date	Aug 28, 2018
Grant date	Aug 28, 2018

Abstract

The present invention relates to a method for selecting a competent oocyte or a competent embryo by determining the expression level of specific microRNA species in a body fluid or in cumulus cells.

First claim

The invention claimed is:

1. A method for selecting an oocyte that will produce, upon fertilization, a viable embryo with a high implantation rate leading to pregnancy, and then implanting an embryo in a female undergoing in vitro fertilization, said method comprising the steps of:
i) measuring, in a cumulus cell surrounding said oocyte, an expression level of at least 5 microRNA selected from the group consisting of hsa-mir-103-1, hsa-mir-103-2, hsa-mir-1826, hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7f, hsa-mir-1244, hsa-mir-182, hsa-mir-21, hsa-mir-30a, hsa-mir-30d, hsa-mir-320a, hsa-mir-508, hsa-mir-92a-1, hsa-mir-92a-2, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-1974, hsa-mir-146b, hsa-mir-886, hsa-mir-210, hsa-mir-1979, hsa-mir-125a, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21;
ii) comparing the expression level of each of said at least 5 microRNA measured in step i) with control expression levels of said at least 5 microRNA measured in cumulus cells associated with competent oocytes;
iii) selecting the oocyte when said at least 5 microRNA are not differentially expressed when compared to cumulus cells associated with competent oocytes;
iv) fertilizing the oocyte selected in step iii) in vitro to generate an embryo; and
v) implanting the embryo generated in step iv) in said female.

2. The method according to claim 1 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-508, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-103-1, hsa-mir-103-2, hsa-mir-1974, hsa-mir-1826, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 12.

3. The method according to claim 1 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-508, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-103-1, has-mir-103-2, hsa-mir-1974, hsa-mir-1826, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 12, hsa-let-7b, hsa-let-7c, hsa-mir-182, hsa-mir-21, hsa-mir-30a and hsa-mir-30d.

4. The method according to claim 1 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-182, hsa-mir-320a, hsa-mir-210, hsa-mir-21 and has-let-7a-1.

5. The method according to claim 1 comprising a further step of measuring in a cumulus cell surrounding said oocyte the expression level of one or more genes selected from the group consisting of ATF3, SIAT6, PRKACA, PLA2G5, GPC6, G0S2, RBMS1, NFIC, SLC40A1 and WNT6.

6. A method for selecting an embryo with a high implantation rate leading to pregnancy, and then implanting said embryo in a female undergoing in vitro fertilization, said method comprising the steps of:
i) measuring, in a cumulus cell surrounding said embryo, an expression level of at least 5 microRNA selected from the group consisting of hsa-mir-103-1, hsa-mir-103-2, hsa-mir-1826, hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7f, hsa-mir-1244, hsa-mir-182, hsa-mir-21, hsa-mir-30a, hsa-mir-30d, hsa-mir-320a, hsa-mir-508, hsa-mir-92a-1, hsa-mir-92a-2, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-1974, hsa-mir-146b, hsa-mir-886, hsa-mir-210, hsa-mir-1979, hsa-mir-125a, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21;
ii) comparing the expression level of each of said at least 5 microRNA measured in step i) with control expression levels of said at least 5 microRNA measured in cumulus cells associated with competent embryos;
iii) selecting the embryo when said at least 5 microRNA are not differentially expressed when compared to cumulus cells associated with competent embryos; and
iv) implanting the embryo selected in step iii) in said female.

7. The method according to claim 6 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-508, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-103-1, has-mir-103-2, hsa-mir-1974, hsa-mir-1826, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 12.

8. The method according to claim 6 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-508, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-103-1, has-mir-103-2, hsa-mir-1974, hsa-mir-1826, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 12, hsa-let-7b, hsa-let-7c, hsa-mir-182, hsa-mir-21, hsa-mir-30a and hsa-mir-30d.

9. The method according to claim 6 wherein said at least 5 microRNA are selected from the group consisting of hsa-mir-182, hsa-mir-320a, hsa-mir-210, hsa-mir-21 and has-let-7a-1.

10. The method according to claim 6 comprising a further step of measuring in a cumulus cell surrounding said embryo the expression level of one or more genes selected from the group consisting of ATF3, SIAT6, PRKACA, PLA2G5, GPC6, G0S2, RBMS1, NFIC, SLC40A1 and WNT6.

11. A method for selecting an oocyte that will produce, upon fertilization, a viable embryo with a high implantation rate leading to pregnancy or an embryo with a high implantation rate leading to pregnancy, and then implanting an embryo in a female undergoing in vitro fertilization, said method comprising the steps of:
i) measuring, in the culture medium of cumulus cell having surrounded said oocyte or said embryo, an expression level of at least 5 microRNA selected from the group consisting of hsa-mir-103-1, hsa-mir-103-2, hsa-mir-1826, hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7f, hsa-mir-1244, hsa-mir-182, hsa-mir-21, hsa-mir-30a, hsa-mir-30d, hsa-mir-320a, hsa-mir-508, hsa-mir-92a-1, hsa-mir-92a-2, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-1974, hsa-mir-146b, hsa-mir-886, hsa-mir-210, hsa-mir-1979, hsa-mir-125a, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21;
ii) comparing the expression level of each of said at least 5 microRNA measured in step i) with control expression levels of said at least 5 microRNA measured in cumulus cells associated with competent oocytes or embryos;
iii) selecting the oocyte or embryo when said at least 5 microRNA are not differentially expressed when compared to cumulus cells associated with competent oocytes or embryos; and
iv) implanting the embryo selected in step iii) in said female or fertilizing the oocyte selected in step iii) in vitro to generate an embryo and then implanting the embryo thereby generated in said female.

12. A method for selecting an oocyte that will produce, upon fertilization, a viable embryo with a high implantation rate leading to pregnancy or an embryo with a high implantation rate leading to pregnancy, and then implanting an embryo in a female undergoing in vitro fertilization, said method comprising the steps of:
i) measuring, in a bodily fluid sample obtained from said female, the expression level of at least 5 microRNA selected from the group consisting of hsa-mir-103-1, hsa-mir-103-2, hsa-mir-1826, hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7f, hsa-mir-1244, hsa-mir-182, hsa-mir-21, hsa-mir-30a, hsa-mir-30d, hsa-mir-320a, hsa-mir-508, hsa-mir-92a-1, hsa-mir-92a-2, hsa-mir-16-1, hsa-mir-16-2, hsa-mir-1974, hsa-mir-146b, hsa-mir-886, hsa-mir-210, hsa-mir-1979, hsa-mir-125a, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO:

Assignees

• Hamamah Samir
• Assou Said
• Inst Nat Sante Rech Med
• Univ De Montpellier 1
• Ct Hospitalier Universitaire Montpellier

Inventors

• Hamamah Samir
• Assou Said

Classifications

• C12Q1/6881 Primary

  for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12Q2600/158

  Expression markers · CPC title

• C12Q2600/178

  miRNA, siRNA or ncRNA · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title