Martinez Rodero, Iris Miralles Bover, Hector

CUMULUS CELLS GENE EXPRESSION AS BIOMARKER FOR

THE ASSESMENT OF DIFFERENT HORMONAL TREATMENTS
IN WOMEN

22nd November, 2016 http://human-fertility.com/wp-content/uploads/2016/06/Fotolia_83587514_S.jpg

 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
 1NDEX
INTRODUCTION

CUMULUS CELLS (CCs) TRANSCRIPTOMICS

CONTROLLED OVARIAN STIMULATION (COS) TREATMENTS

CCs AS BIOMARKERS FOR THE DIFFERENT COS TREATMENTS

METHODOLOGICAL FRAMEWORK FOR THE IMPLEMENTATION

CONCLUSIONS
Figure 1. Female reproductive organs representation
(http://www.hormone.org/~/media/hormone/images.jpg)
1. INTRODUCTION
 ARTs context

Figure 5. ARTs social landscape

(http://www.safefertilitycenter.com)
ARTs context
 ARTs context

Figure 2. Embryo transference in women

(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

 ARTs context

avoid multiple pregancies

Figure 2. Embryo transference in women
(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

Figure 3. Double pregnancy in women

(https://encrypted-tbn0.gstatic.com/images)
 ARTs context

avoid multiple pregancies

Figure 2. Embryo transference in women
(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

Figure 3. Double pregnancy in women

(https://encrypted-tbn0.gstatic.com/images)
 Figure 4 . 8 cell embryos of diferent quality, classified in A, B and
C (https://invitra.com//Embryo-classification470x200.jpg)

ARTs context

to select THE BEST EMBRYO

avoid multiple pregancies

Figure 2. Embryo transference in women
(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

Figure 3. Double pregnancy in women

(https://encrypted-tbn0.gstatic.com/images)
 Figure 4 . 8 cell embryos of diferent quality, classified in A, B and
C (https://invitra.com//Embryo-classification470x200.jpg)

ARTs context

to select THE BEST EMBRYO

avoid multiple pregancies

Figure 2. Embryo transference in women
(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

Figure 3. Double pregnancy in women

(https://encrypted-tbn0.gstatic.com/images)
 Figure 4 . 8 cell embryos of diferent quality, classified in A, B and
C (https://invitra.com//Embryo-classification470x200.jpg)

ARTs context

Cumulus Cells (CCs) to select THE BEST EMBRYO

avoid multiple pregancies

Figure 2. Embryo transference in women
Genetic Profiling
(http://www.fwivf.com/images/photos/home-
mast.png)

single embryo transference

Figure 3. Double pregnancy in women

(https://encrypted-tbn0.gstatic.com/images)
 Steps in IVF

ARTs context
 Steps in IVF

ARTs context

Figure 6. Steps in IVF treatment (https://encrypted-tbn3.gstatic.com/images)

ARTs context
ARTs context
ARTs context

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
ARTs context

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
ARTs context

to understand
molecular
mechanisms

to predict
treatments
outcome

CUMULUS CELLS BIOMARKERS

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
ARTs context

to understand
molecular
mechanisms

to predict
treatments
outcome

CUMULUS CELLS BIOMARKERS

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
ARTs context

to understand
molecular
mechanisms

to predict
treatments
outcome

CUMULUS CELLS BIOMARKERS

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
ARTs context
to understand
molecular
mechanisms

to predict
treatments
outcome

CUMULUS CELLS
BIOMARKERS

Figure 7. Doctor ofering different treatments

(http://healthaffairs.org/blog/wp-content/uploads.jpg)
2. CUMULUS CELLS TRANSCRIPTOMICS
Why can we use CCs transcriptomics to study oocyte quality?
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 8. Follicles in development and their relationship with the oocyte (Guzeloglu-Kayisli et al. Biochemical Journal 2012;446:47–58 (10))
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 8. Follicles in development and their relationship with the oocyte (Guzeloglu-Kayisli et al. Biochemical Journal 2012;446:47–58 (10))

CUMULUS
CELLS
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 8. Follicles in development and their relationship with the oocyte (Guzeloglu-Kayisli et al. Biochemical Journal 2012;446:47–58 (10))

CUMULUS ensurement of oocyte correct development

CELLS
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 8. Follicles in development and their relationship with the oocyte (Guzeloglu-Kayisli et al. Biochemical Journal 2012;446:47–58 (10))

CUMULUS ensurement of oocyte correct development

CELLS
bidirectional communication
Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing
 Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 9. Oocyte gene expression (Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways.)
 Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 9. Oocyte gene expression (Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways.)
 Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

Figure 9. Oocyte gene expression (Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways.)
 Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

expression of
specific genes

Figure 9. Oocyte gene expression (Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways.)
 Why can we use CCs transcriptomics to study oocyte quality?
Tight relationship between oocyte and cumulus cells during growing

expression of
specific genes

morphological
and
physiological
characteristics
of the oocyte

Figure 9. Oocyte gene expression (Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways.)
Technical issues:
Technical issues: qrtPCR + microarraysç
Technical issues: qrtPCR + microarraysç
Technical issues: qrtPCR + microarrays
 Technical issues: qrtPCR + microarrays

1. SINGLE GENE ANALYSIS

 Technical issues: qrtPCR + microarrays

1. SINGLE GENE ANALYSIS

Figure 10. Polimerase Chain Reaction (PCR) squeme

(http://ib.bioninja.com.au/_Media/pcrcomponents_med.jpeg)
 Technical issues: qrtPCR + microarrays

1. SINGLE GENE ANALYSIS 2. MULTIPLE GENE ANALYSIS

Figure 10. Polimerase Chain Reaction (PCR) squeme

(http://ib.bioninja.com.au/_Media/pcrcomponents_med.jpeg)
 Technical issues: qrtPCR + microarrays

1. SINGLE GENE ANALYSIS 2. MULTIPLE GENE ANALYSIS

Figure 11. Microarray squeme

Figure 10. Polimerase Chain Reaction (PCR) squeme (http://image.slidesharecdn.com/biotechnology-microarray)
(http://ib.bioninja.com.au/_Media/pcrcomponents_med.jpeg)
 Technical issues: qrtPCR + microarrays

1. SINGLE GENE ANALYSIS 2. MULTIPLE GENE ANALYSIS

GENE EXPRESSION

VARIES BETWEEN

SUPEROVULATION TREATMENTS

Figure 10. Polimerase Chain Reaction (PCR) squeme Figure 11. Microarray squeme
(http://ib.bioninja.com.au/_Media/pcrcomponents_med.jpeg) (http://image.slidesharecdn.com/biotechnology-microarray)
3. CONTROLLED OVARIAN STIMULATION
TREATMENTS
 (GnRh)
Controlled Ovarian Stimulation
hipofisis
 Avoid early follicular luteinisation

Figure 12. Types of GnRH analogues (Maria Jesús Barros San

Cristóbal (2014). Agonistas versus antagonistas: resultados
en FIV e ICSI. Universidad de Oviedo )

same oocyte quality

differences when using agonists and antagonists
same CCs gene expression
 Follicular Recruitment

HMG FSH

LH and FSH FSH activity

activity

Multiple
Recruit and folicular
helps development
maturation

Figure 13. Hormonal levels compared in normal and estimulated cicle (Lizán Tudela, C. (2012). Efectos de
la estimulación ovárica en la calidad embrionaria. Universitat de València)

different oocyte quality

different
different effects when using FSH or hMG
IVF outcome
different CCs gene expression
 Ovulation triggering

LH and hCG bind

the same receptor

LH and hCG share:

the same α subunit

the 81% of the Aa of the β subunit

Figure 14. Structure of LH and hCG receptors (Slideshare.net/sandroesteves/v2-rec-hcg-in-

artmerksorono-symposiumsingapore2014)
3. PREDICTIVE POTENCIAL OF CCs MARKERS
FOR THE DIFFERENT COS PROTOCOLS
 Different pharmacodinamics
FSH
Change in follicle gene expression
HMG Disrupt celular events respect to
their normal activity

Figure 15. Esteroidogenesis during a natural cycle

(Requena Miranda, A. (2013) Universidad Rey Juan Carlos)

Figure 16. Internal factors affecting cells

(Gomez Urviola, Slideshare (2009): es.slideshare.net/gomezurviola/biocelmol-16)
 Predictive potencial of CCs markers

Genotype + Environment (hormone treatment) = PHENOTYPE

differential patterns in cumulus cells gene expression between

FSH and hMG treatments allows the identification of a tissue or
cell phenotype

EVALUATION OF EMBRYO QUALITY

from ocytes obtained by either one treatment or another is
possibe thanks to CCs markers

Figure 17. PCA analysis of Cumulus Cells and differentiatin between genetic expression
pattens. H: hMG stimulated cycle, F: uFSH stimulated cycles, G: rFSH stimulated cycles
((Requena Miranda, A. (2013) Universidad Rey Juan Carlos)
 4. METHODOLOGICAL FRAMEWORK
FOR CCs MARKERS IMPLEMENTATION
CONCLUSIONS
CONCLUSIONS
CONCLUSIONS
CONCLUSIONS
WHATEVER WILL HAPPEN WITH
CUMULUS CELL MARKERS, THEY ARE
THE FUTURE....
WHATEVER WILL HAPPEN WITH
CUMULUS CELL MARKERS, THEY ARE
THE FUTURE....

LETS TAKE IT!

THANK YOU FOR YOUR
ATTENTION
Martinez Rodero, Iris Miralles Bover, Hector

CUMULUS CELLS GENE EXPRESSION AS BIOMARKER FOR

THE ASSESMENT OF DIFFERENT HORMONAL TREATMENTS
IN WOMEN

22nd November, 2016 http://human-fertility.com/wp-content/uploads/2016/06/Fotolia_83587514_S.jpg

 Technical issues: quantitative real time PCR (qrtPCR)

Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways. Translocations, 45, 46XY.
 Technical issues: Microarrays

Matzuk, M. M., & Lamb, D. J. (2002). Genetic dissection of mammalian fertility pathways. Translocations, 45, 46XY.