Serono Research Grants, 2005
Field of Clinical Research in Endocrinology
Research Project:
Relationship between cannabinoid receptors and fatty acid composition in morbid obesity.
Main researcher:
Dr. Eduardo García Fuentes.
Project Summary:
The prevalence of obesity has increased considerably in recent years, becoming recognized as a problem with epidemic characteristics. Obesity is the result of a complex metabolic disorder in which there is an extended imbalance between energy intake and total energy expense.
The association between obesity and insulin resistance is well documented. It seems that fatty acids play a relevant role in said relationship, although the mechanisms are not well known. Fatty acids are not just a source of energy. They form part of numerous lipid complexes, of membranes and they act as potent metabolic intermediate products, the type of fatty acid being very important in the effect produced. These fatty acids influence the expression of certain transcription factors (including SREBP-1c) and they can modify the activity of insulin-activated intracellular pathways (phosphatidylinositol 3-kinase –PI3K- and mitogen-activated protein kinase -MAPK-).
These two kinases are also stimulated by cannabinoid receptor activation. Regulation of these receptors, specifically CB1, is important in consumption and appetite control. Despite the central nervous system being the main location for endocannabinoid action, there is evidence indicating that these molecules also exert their actions upon the adipose tissue, regulating lipid and hydrocarbonate metabolism, at least in animals.
This project intends to further our knowledge in the relationship existing between food intake control by the cannabinoid system, the state of insulin resistance and the influence that the kind of fatty acids can exert on these, in a patient model (morbid obese) in which there is a hydrocarbonate and lipid metabolism disorder.
Jury:
Field of Clinical Research in Infertility
Research Project:
Functional characterization of lymphocyte subpopulations in women with repeat abortions by primary syndrome of antiphospholipid antibodies.
Main researcher:
Dr. Javier Carbone Campoverde.
Project Summary:
The presence of recurrent abortion constitutes a potentially frustrating problem both for the women suffering them and for the doctors treating them. The Primary Antiphospholipid Antibody Syndrome (PAPS) is the first cause of abortion due to immunological disease and one of the few treatable causes. However, the abortion mechanism in PAPS is still not well known. Aim: To characterize alterations of functional lymphocyte subpopulations in women with obstetric PAPS and whether there is an association between these alterations with clinical and immunological activity parameters for obstetric PAPS. Methodology: Analytical crosswise observational study. Patients: 36 women with repeat abortions with antiphospholipid antibodies (APA). Control Groups: 36 women with recurrent abortions not presenting APA or any known cause for abortions, 36 women who have had children with no abortions and 36 women with positive APA without PAPS symptoms. The patients will be selected in External Practices of Clinical Immunology, Obstetrics and Infertility and Genetics of the General University Hospital Gregorio Marañón. Presence of APA: ELISA technique for IgG and IgM isotype APA dependent on Beta-2-glycoprotein-I and coagulometric techniques with topic anticoagulant. Quantification of functional lymphocyte subpopulations by direct immunofluorescence and analytical flow cytometry. Panel of monoclonal antibodies: CD38 FITC/ HLADR PE /CD4 or CD8PerCP; CD40 FITC/CD5 PE/CD19 PercP; CD25 FITC/CD45RO PE/CD4 PerCP; CD56 FITC/CD69 PE/CD3 PercP and isotopic controls. A substudy will be performed in order to establish the immunophenotype of T cells activated after stimulation with beta-2-glycoprotein-I by means of staining with CFSE. Statistics: variable normality (Kolmogorov-Smirnov); mean comparison (ANOVA or Kruskall Wallis); association between variables (Pearson or Spearman); multiple linear regression analysis in order to determine CD4+ and CD8+ T cell subpopulations most strongly associated with the presence of APA.
Jury:
Field of Multiple Sclerosis Clinical Research
Research Project:
Prognostic markers in patients with clinically isolated syndrome. Conversion in defined Multiple Sclerosis and disease progression.
Main researcher:
Dr. Rafael Arroyo González
Project Summary:
The objectives of the present study are:
1. The search for molecular markers directly related to the Inflammation-Demyelination-Axonal damage processes in Multiple Sclerosis (MS) that would allow predicting the conversion of a clinically isolated syndrome (CIS) into clinically defined MS; to this end we will calculate the correlation between the markers of:
a. Inflammation (TNF alpha, IL-10, IL-12) and gadolinium uptake by magnetic resonance (MR);
b. Axonal damage (Tau and 14-3-3) and total NAA peak in MR Spectroscopy;
c. Antibody-mediated demyelinization (anti-MOG and anti-MBP) and lesion number in T2 and gadolinium uptake, and total NAA peak in MR and MR Spectroscopy, respectively;
d. Inflammation and axonal damage;
e. Inflammation and antibody-mediated demyelinization;
f. Antibody-mediated demyelinization and axonal damage.
2. To predict MS evaluation by means of patient follow-up during a period not under two years, in which we will compare the values obtained for inflammation markers, demyelinization and axonal damage with the number of outbreaks, outbreak seriousness and worsening of EDSS greater than 1 point (if EDSS<5.5) or greater than 0.5 points (if EDSS>=5.5).
In order to meet these aims, an observational study of cases (patients with clinically defined MS from CIS) and controls (CIS patients with findings suggesting an etiology different to MS) in which no less than 30 patients will be included in each group and who will be followed up during a period not less than 2 years (with basal visit and visits 3, 6, 12, 18 and 24 months from then, and a visit in case of an outbreak). Samples of cerebrospinal fluid (CSF), blood and serum will be taken during the basal visit, and only the latter two during the remaining visits; in the basal visit and in the 12 and 24 month visits conventional and spectroscopic MR will be performed. mRNA will be extracted from the blood samples and the expression of genes coding for TNF alpha, IL-10 and IL-12 will be measured by real time quantitative RT-PCR. The Tau and 14-3-3 proteins as well as the anti-MOG and anti-MBP antibodies will be quantified by ELISA from the CSF and serum.
Jury:
Field of Psoriasis Clinical Research: Clinical Analysis and Inmunology
Research Project:
A study of the gene expression profile as a marker for therapeutic response to different biological drugs in systemic treatment of Psoriasis.
Main researcher:
Dr. Esteban Daudén Tello
Project Summary:
One of the main problems facing clinical pharmacology is the large interindividual variability existing in the response to drugs, both regarding effectiveness and toxicity, such that different patients respond differently to the same medication. This is due to genetic and non-genetic factors. Gene expression, more than the genetic composition itself, and the polymorphisms existing in the genes is what explains and determines, at least in part, these differences. The research of variables that may allow us to determine the degree of response prior to beginning treatment with a pharmacological agent must be a prime objective in current medicine. The field of pharmacogenetics has opened a new horizon in this direction and psoriasis should not be different to this situation. However, although in recent years studies have been performed in very different diseases with different pharmacological agents, psoriasis has been left aside. We thus consider justified carrying out the present Research Project that would undoubtedly contribute great benefits to optimizing the therapeutics of this complex disease.
Aims of the study
Main Aim
• To determine the gene expression profile before and after systemic administration of different biological drugs in patients with chronic psoriasis in moderate-severe plaques and establish a correlation with therapeutic response such as to determine a priori the efficacy and tolerance of drugs for each patient and thus select “the correct drug for each patient”.
Secondary Aims
• To increase knowledge on the action mechanism at molecular level of the study drugs.
• To increase knowledge on physiopathological mechanisms involved in psoriasis.
• To identify therapeutic targets that allow research and development of new drugs.
• To create and maintain a gene bank of patients with chronic psoriasis in plaques with samples obtained at basal level (blood and skin tissue) and after 12 weeks of treatment (skin tissue).
• To evaluate effectiveness and safety of the study drugs in chronic psoriasis treatment in moderate-serious plaques during 12 weeks of follow-up.
Study design
This is a prospective study in patients with chronic psoriasis in moderate-severe plaques (PASI greater than or equal to 10 and BSA greater than or equal to 10% of body surface) with or without psoriatic arthritis that do not respond adequately or are not susceptible to topic treatment or phototherapy and with indication in order to perform systemic treatment.
The patients will receive therapy with some of the following biological drugs authorized for indication of psoriasis and/or psoriatic arthritis in monotherapy: efalizumab, etanercept and infliximab (in the case of patients without associated psoriatic arthritis, the drug will be requested by compassionate use). The choice of one drug or another will be according to the researcher’s criteria, as long as it corresponds with the indication of the summary of product characteristics for each product. The dose, administration route and posology regime will also correspond to that indicated on the summary of product characteristics. The patients will receive continuous treatment during at least 12 weeks.
The study includes a basal visit (start of treatment), monthly follow-up visits (4 and 8 weeks after the basal visit) and a final visit (12 weeks after starting treatment). The following psoriasis activity parameters will be assessed: PASI, BSA, sPGA, PGA of change, PGPA, pruritus scale, PsA-GA, DLQI, EuroQol-5D and EuroQol thermometer. Safety will be analyzed by considering the following variables: registering adverse effects, vital constants, physical examination, concomitant medication and supplementary analytical tests. The Morisky-Green test will be used in order to evaluate the degree of therapeutic compliance.
A pharmacogenetic study will be performed in order to evaluate gene expression (RNA study) related to the therapeutic response to the different drugs studied in terms of effectiveness and toxicity. During the basal visit and in the final visit a blood sample and two tissue samples will be obtained by means of skin biopsy (lesional and non-lesional skin). The following procedures will take place in the RNA analysis: RNA extraction from the samples, amplification, sequencing and RNA identification (by means of a high density oligonucleotide microarray for evaluating gene expression) with fluorescence reading through a laser scanner. A bioinformatics analysis (digital image analysis, summarization, normalization and statistical study) will subsequently be performed on the data obtained.
Once expressed genes and their quantitative levels are identified a comparative analysis will be performed between the blood samples between lesional skin and non-lesional skin and this will be correlated with the parameters of psoriatic activity assessment and the appearance of adverse events for each drug. The analysis of these results will allow determining a priori molecular markers for therapeutic response regarding efficacy and tolerance of the drugs for each patient in order to carry out personalized medicine.
Jury:
Bolsa Serono de Investigaçâo Clínica em Psoriase
Research Project:
T-Cell subsets modulation in psoriatic lesions by photo (chemo) therapy and systemic biological agents.
Main researcher:
Dr. Paulo Leal Filipe
Project Summary:
Psoriasis is a skin disorder characterised by an increased proliferation and disturbed differentiation of keratinocytes. Current therapies, photo (chemo) therapy and systemic biological agents for moderate to severe psoriasis can control the disease but are not curative.
The main objective of the study is to characterise the immune pattern and the markers of the inflammatory cascade, hyperproliferation and keratinisation in the psoriatic lesions of patients with moderate to severe plaque psoriasis. The individual response to conventional PUVA and narrow band UVB therapies and systemic immuno-target drugs, such as, etanercept and efaluzimab will be assessed through Psoriasis Area and Severity Index (PASI) and histological indexes based on keratinisation and epidermis hyperplasia reduction.
T-cell subsets for CD2+, CD4+, CD8+, CD11a+, CD25+, CD45RO+, CD45RA+, CD103+ antigens, relevant in the immunopathogenesis of psoriatic lesions, will be assessed at the baseline and at the first evaluation after photo (chemo) therapy or systemic immunomodulation therapy stars. Besides, the correlation of the T-lymphocyte activation profile with the levels of TNFα and markers of kerationocyte hyperproliferation (Ki67) and keratinisation (K10 and K16) at both assessment phases will be investigated. The quantitative histological evaluation of epidermis will enable to establish plaque remission indexes, more accurately than clinic response indexes assessed through PASI, and a T-cell profile in responders and non-responders. Also, the skin barrier function will b e evaluated inspecting the morphological alterations and calcium and other divalent ions involvement in epidermis remodelling. The study will contribute to a better knowledge of the immune response in the psoriasis lesion and to the characterisation of the immunological T-cell profile in different therapeutic approaches.