Charles University

Description of the legal entity:

Charles University (CUNI) was founded in 1348, making it one of the oldest universities in the world. It is the largest and most renowned Czech university, and is also the best-rated Czech university according to international rankings. The key priority of Charles University is to continue to enhance its prestigious status as a research university. To achieve this aim, the University focuses strongly on research activities.

Laboratory of Pathophysiology of Adipose Tissue belongs to the Department of Pathophysiology, Third Faculty of Medicine, Charles University continues and expands the research done in former Department of Sport Medicine. The Laboratory presented a number of important contributions in studies focused at changes of metabolic, endocrine and immune characteristics of AT during lifestyle –diet and physical activity- interventions in obese population. These studies have been performed frequently in collaboration with French laboratory of Obesity research of INSERM in Toulouse and, also, in the frame of several European projects supported by European Commission. The main aim of the present activity is to elucidate a role of lipogenic and adipogenic capacity and inflammatory state of AT in ethiopathogenesis of obesity, type 2 diabetes, lymphedema and in pathogenesis of metabolic disturbances associated with aging. Laboratory is focused on clinical and translational research combining clinical studies with patients and in vitro laboratory techniques to analyze processes that underlie, or mediate disturbances related to dysfunctional adipose tissue.

In TheraLymph, the Laboratory will be dedicated to elucidation of the crosstalk between LEC and adipocytes. The successful accomplishment of this task is based on our extensive experience with in vitro cultivation and differentiation of adipocytes, on the available cryobank of preadipocytes derived from various subjects including subjects with lymphedema, experience with angiogenic assays using HDMEC, HUVEC and adipose tissue explants. Moreover, we currently develop protocols for coculture of HDMEC and preadipocytes that can be easily translated to HDLEC.

Curriculum Vitae:

Lenka Rossmeislova, PhD (Female): a head of Laboratory of Pathophysiology of Adipose Tissue (belonging to the Department of Pathophysiology, Third Faculty of Medicine, Charles University). She is a scientist with 20 years of expertise on adipose tissue. Besides expertise in adipose cell biology (characterization of  human adipose tissue and its cellular components, study of metabolism  and secretion products of adipose tissue, analysis of stress of endoplasmic reticulum in adipose tissue, obesity, hypocaloric diets, cellular senescence, study of the role of lymphatic stasis and interstitial fluid in adipose tissue expansion), she has substantial experience with confocal microscopy and imaging techniques. Her recent research grant was dedicated to the elucidation of lymphatic stasis as a modulator of adipose tissue expansion and inflammation.

H index 15, 32 papers in IF journals, number of citations 1002.

She will coordinate all experiments done by CUNI partner.

Michal Koc, PhD (Male): a senior scientist of Laboratory of Pathophysiology of Adipose Tissue. He is a molecular biologist with 20 years of laboratory experience with deep knowledge of diverse molecular biology techniques including shRNA technology and lentiviral transduction.

H index 11, 20 papers in IF journals, number of citations 345.

Zuzana Varaliova (Female):  Zuzana is a PhD student led by Dr. Rossmeislova. She has been trained in cell cultures of both adipocytes and endothelial cells and assessment of gene expression.

Medical doctors, nurses and technical personnel will be determined later. These team members will enable collection and analysis of adipose tissue from various subjects. Accountant will be responsible for the budget management and reporting.

CUNI Main tasks in the project per WP

• WP3: Adipose tissue function in lymphedema (Task 3.1), Effect of FFA on lymphatic function (Task 3.2)

WP9 CUNI will contribute to dissemination and communication activities

List of up to 5 relevant publications, and/or products, services (including widely-used datasets or software), or other achievements relevant to the  call content

• Cucchi F, Rossmeislova L, Simonsen L, Jensen MR, Bulow J. (2017) A vicious circle in chronic lymphoedema pathophysiology? An adipocentric view. Obes Rev 18, 1159-1169.
• Rossmeislova, L., L. Malisova, J. Kracmerova, M. Tencerova, Z. Kovacova, M. Koc, M. Siklova-Vitkova, N. Viquerie, D. Langin and V. Stich (2013). “Weight Loss Improves the Adipogenic Capacity of Human Preadipocytes and Modulates Their Secretory Profile.” Diabetes 62(6): 1990-1995.
• Janderova L, McNeil M, Murrell AN, Mynatt RL, Smith SR. Human mesenchymal stem cells as an in vitro model for human adipogenesis. Obesity Research. 2003;11:65-74
• Kovacikova M, Sengenes C, Kovacova Z, Siklova-Vitkova M, Klimcakova E, Polak J, Rossmeislova L, Bajzova M, Hejnova J, Hnevkovska Z, Bouloumie A, Langin D, Stich V. Dietary intervention-induced weight loss decreases macrophage content in adipose tissue of obese women. Int J Obes (Lond). 2011;35:91-98

Siklova-Vitkova M, Klimcakova E, Polak J, Kovacova Z, Tencerova M, Rossmeislova L, Bajzova M, Langin D, Stich V. Adipose tissue secretion and expression of adipocyte-produced and stromavascular fraction-produced adipokines vary during multiple phases of weight-reducing dietary intervention in obese women. J Clin Endocrinol Metab. 2012;97:E1176-1181

List of up to 5 relevant previous projects or activities, connected to the subject of this proposal

Grant Agency of the Czech Republic 2016-2018, “LYMPHAT- Lymphatic stasis as a modulator of adipose tissue expansion and inflammation”.

Infrastructure and/or major items of technical equipment

Describe any significant infrastructure and/or any major items of technical equipment, relevant to the proposed work.

Facilities necessary and available to the project include biohazard cell culture hood, incubator, inverted microscope Nikon TS100F equipped with digital camera,  laboratory space equipped for DNA and RNA isolation and analysis (Nanodrop spectrophotometer, PCR cycler), Western immunoblotting (BioRad Miniprotean Tetra), ELISA (microplate reader Versamax with monochromator) and general laboratory function. Partner has an access to ABI PRISM 7500 Real Time RT-PCR equipment Leica TCS SP5 confocal microscope, XCellIgence and KODAK ImageStation.

Operational capacity

Fully operational