Serono Research Grants, 2003

Field of Clinical Research in Endocrinology and Metabolism

Research Project:

Combined treatment with Flutamide-Metformin in teenagers with type 1 Diabetes and ovarian hyperandrogenism: effects on metabolic control, ovarian function and corporal composition.

Main researcher:

Dra. Lourdes Ibáñez Toda

Project Summary:

Adolescents with type 1 diabetes show deterioration of metabolic control during puberty, largely due to an increase in insulin needs, which can determine the development of peripheral hyperinsulinism and an excessive increase in weight. As well as favoring the precocious appearance of microvascular complications, hyperinsulinism can condition the appearance of menstrual disorders, an increase in serum concentrations of androgens and development of infertility.

In non-diabetic women, hyperinsulinism is a key etiopathogenic factor in developing ovarian and suprarenal hyperandroginism. Population differences in the prevalence of this disturbance seem to be also related to the length of the CAG sequence that codifies the androgen receptor. In women with ovarian hyperandrogenism associated with hyperinsulinism the administration of insulin sensitizing agents (metformin) determines a reduction in insulin and androgen figures and regularizes menstruation cycles making them ovulatory. The combined administration of metformin and antiandrogens (flutamide) is even more effective upon the variables mentioned, and also determines favorable changes in body composition, specifically a reduction in total and abdominal fat mass, the excess of which is considered a factor of cardiovascular risk.

In diabetic women, preliminary studies seem to demonstrate that the administration of metformin as a coadyuvant of insulinotherapy reduces the needs for insulin and favors maintaining body weight. However, it is unknown whether extended administration of this insulin sensitizer, combined with an antiandrogenic drug in adolescents with type 1 diabetes can prevent or reverse the negative effects of pubertal insulin resistance on ovarian and suprarenal function, as well as on body composition.

The objectives of the study are: 1) To determine the prevalence of ovarian hyperandrogenism in adolescents with type 1 diabetes; 2) To establish the relationship between the presence and seriousness of ovarian hyperandrogenism, metabolic control, cardiovascular risk factors and the length of the androgen CAG receptor sequence; 3) To determine whether the combined treatment with flutamide and metformin is capable of reversing the clinical manifestations of ovarian hyperandrogenism, alterations of corporal composition, reducing the levels of androgens, restoring ovulation and improving metabolic control.

Methodology: Prospective, randomized, open and controlled study. Subjects and scope of study: 100 adolescents with type 1 diabetes (First phase) controlled in the Endocrinology departments of two hospitals in the area of Barcelona; in a second phase, the patients in which the existence of ovarian hyperandrogenism (hirsutism, menstrual disorders, an increase in serum androgens and chronic anovulation) is identified will be studied.
Implementation: First phase: to determine the presence of ovarian hyperandrogenism. Second phase: ovulation function will be studied in these patients for 3 months, and they will then randomly receive therapy with metformin (1275 mg/d) and flutamide (62,5 mg/d) or they will remain without treatment for 9 months.
Determinations: 1) Clinical variables: weight, stature, body mass index, waist-hip ratio, degree of hirsutism; 2) Analytical variables: first and second phase: HbA1c, microalbuminuria, androgens, LH, FSH, estradiol, SHBG, lipid profile; second phase: ovulatory function (weekly determination of progesterone on filter paper); 3) Body composition: measurement of total fat mass and specific body regions (torso and abdomen); clinical and analytical variables will be studied before and after the treatment or control period; 4) Genetic study: study of the CAG sequence of the androgen receptor.

Analysis:
comparison between endocrine-metabolic parameters: Student’s t test; genotype/endocrine-metabolic variable ratio: ANOVA; genotype/clinical and analytical variable ratio: multiple regression analysis.

Jury:

Field of Assisted Human Reproduction

Research Project:

Application of cytochemical and ultrastructural techniques on human gametes for clinical study of fertility failure (and anomalous fertility) as a cause for sterility in human assisted reproduction. Deeper understanding of molecular study of interaction between human gametes.

Main researcher:

Dr. Pedro J. Fernández-Colom

Project Summary:

The fecundity process in mammals requires an ordered series of events among which is the recognition between complementary receptors and ligands located in the sperm membranes and pellucid area, as well as the subsequent fusion of gametes. Clinical practice in assisted reproduction techniques shows that, occasionally, the in vitro fertilization process does not occur or it takes place abnormally. Many of the clinical cases in fertility failure cannot be explained, and therefore it is impossible to perform a correct diagnosis and treatment of sterility. The present project intends to approach this subject by setting the main aims of:
1. Studying the composition of glycoproteins of pellucid area of human oocytes potentially involved in the interaction with the human spermatozoid.
2. To deepen in the knowledge of the fertility process in the human species by studying molecular changes produced in the pellucid area during oocyte maturation (zonal maturation) and after releasing the content of cortical granules (zone reaction).
3. To identify the presence and distribution of receptors presumably implied in the fertilization process that are present in the plasmatic and acrosomal membrane of human spermatozoids. To determine how in vitro selection of spermatozoids for assisted reproduction techniques may affect said distribution.
4. To diagnose the possible existence of molecular pathologies in human gametes as a cause of fertility failure or anomalous fertilization phenomena in the scope of Human Reproductive Medicine by means of the combined use of different technologies (immunohistochemistry, ultrastructure, hemizona test, acrosomal reaction test, etc.).

Jury:

Field of Multiple Sclerosis Clinical Research

Research Project:

Implication of dendritic cells in precocious multiple sclerosis physiopathology. Development of an in vitro model of tolerance induction with dendritic cells.

Main researcher:

Dra. Silvia Sánchez Ramón

Project Summary:

The present project consists in a prospective observational study in a cohort of 40 patients with clinical start of multiple sclerosis (MS) in collaboration between the Immunology and Neurology Department of the Gregorio Marañón General University Hospital. The role of dendritic cells (DCs) and regulatory T cells (Treg) in precocious MS physiopathology when neurological deficits are still reversible. We will study the number, phenotype and function of DC and Treg populations present in the cerebrospinal liquid (CSL) with multiparametric flow cytometry and direct immunofluorescence and with respect to a Treg in peripheral blood in two control groups of healthy subjects and patients with rheumatoid arthritis as a paradigm of Th1 autoimmune disease. Secondly, we will study the functional capacity of isolated DCs of CSL by means of lymphoproliferative studies, flow cytometry and enzymoimmunoanalysis. The data obtained will be correlated with the clinical and paraclinical study variables and with disease progression.

Likewise, given the relevant role of DCs in immunological response regulation, we will develop a tolerance model for T lymphocytes against myelin proteins by means of DC manipulation. After selecting and purifying or generating immature peripheral blood DCs from patients with MS, these will be exposed to MS specific antigens and the role of modulatory molecules (costimulators and cytokines) in T lymphocyte polarization in a DC:Lymphocyte T coculture model.

Jury:

Field of Psoriasis Clinical Research: Clinical Analysis and Immunology

Research Project:

Implication of CLA+ T lymphocytes with skin tropism in psoriasis physiopathology in acute outbreak and chronic evolution plaques.

Main researcher:

Dr. Ramón María Pujol Vallverdú

Project Summary:

CLA+ T lymphocytes are cells that are capable of inducing inflammation in numerous skin pathologies mediated by T cells such as atopic or contact eczema. The aim of this project is to study the role of CLA+ T lymphocytes as the starters of inflammatory response in psoriasis, in keratinocyte activation and in maintaining chronic inflammation through its effects on the fibroblasts. For this, peripheral blood and skin biopsies would be obtained from patients with psoriasis in acute and chronic plaques, as well as in healthy controls. The phenotype of activated and resting T lymphocytes will be typified valuing its capacity for activating keratinocytes and fibroblasts. Among other mediators, the production of IL-20, a cytosine involved in keratinocyte proliferation by CLA+ T cells, will be determined. Induction of IL-7 will be studied by keratinocytes and fibroblasts, CLA+ and CLA- circulatory memory highly purified with detection by different mediators by Real Time RT-PCR. In all cases the control will be the CLA- memory T cells of the same individual. The most relevant mediators produced selectively by these cells will be confirmed in skin biopsies by means of Real Time RT-PCR and/or in culture cells by ELISA.

Jury: