- Email: [email protected]
A role for keratinocytes in developmental myelopoiesis
New Biotechnology · Volume 33, Number 3 · May 2016
POSTERS
PS – R11
2. New therapeutic and diagnostic strategies A role for keratinocytes in developmental myelopoiesis Francisco Juan Martínez-Navarro ∗ , Diana García-Moreno, Azucena López Mu˜noz, José Meseguer, Victoriano Mulero Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
E-mail address: [email protected] (F.J. Martínez-Navarro). Tnfa is a powerful pro-inflammatory cytokine which signals through two receptors, namely Tnfr1 and Tnfr2. Tnfr2 has been shown to protect skin from oxidative stress-induced inflammation using the zebrafish as an experimental model. In the present study, using fluorescent reporter transgene, fluorescence microscopy and RT-qPCR, we studied the impact of Tnfr2 overexpression in zebrafish keratinocytes. Surprisingly, overexpression of Tnfr2 in keratinocytes led to increase neutrophil numbers in 3 days postfertilization larvae concomitantly with decreased transcript levels of Tnfa and increased levels of Il1b, Pu.1, Gata1, Mcsf and Gcsf. These results suggest a critical importance of keratinocytes in developmental myelopoiesis and may explain the systemic neutrophilia associated to several skin inflammatory disorders, such as the acute febrile neutrophilic dermatosis or Sweet’s syndrome.
Following this strategy, we have previously reported how a new collection of 4-alkoxy-6,9-dichloro[1,2,4]triazolo[4,3a]quinoxalines exhibit potent in vitro anti-inflammatory activity as inhibitors of the pro-inflammatory cytokines IL-6 and TNF-␣. Our main aim now is to study how these quinoxaline derivatives act upon different intracellular signaling mechanisms, in an attempt to find their potential molecular targets. We have identified a group of intracellular molecules whose patterns of activation and/or expression are altered by the analyzed compounds, providing further information about the inhibition of TNF-␣ and IL-6. Our results may be of great interest for the pharmaceutical industry, as a starting point for the development of new and more potent anti-inflammatory drugs derived from quinoxalines. http://dx.doi.org/10.1016/j.nbt.2015.10.018
PS – R13
3. Bioenergy and bioremediation Synthetic biology and metabolic engineering to produce microbial oils: towards a sustainable source of fuels, chemicals and nutraceuticals Rodrigo Ledesma-Amaro 1,∗ , Jose Luis Revuelta 2 1
2
INRA, AgroParisTech, UMR1319 Micalis, F-78350 Jouy-en-Josas, France Universidad de Salamanca, Salamanca, Spain
http://dx.doi.org/10.1016/j.nbt.2015.10.017
PS – R12
2. New therapeutic and diagnostic strategies Discovering the molecular targets of 4-alkoxy-6,9dichloro[1,2,4]triazolo[4,3-a]quinoxalines that mediate an anti-inflammatory effect on the macrophage function María Tristán-Manzano 1,∗ , Antonio Guirado 2 , María MartínezEsparza 1 , Pilar García-Pe˜narrubia 1 , Antonio J. Ruiz-Alcaraz 1 1
Department of Biochemistry, Molecular Biology and Immunology B, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain 2 Department of Organic Chemistry, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain
E-mail address: [email protected] (M. Tristán-Manzano). A dysregulation of the inflammatory process, which is a complex biological response directed by the immune system to maintain homeostasis, leads to the development of chronic inflammatory or autoimmune diseases, whose current treatments present many side effects. Nowadays, the search of new therapeutic alternatives focuses on different strategies. Some of them are orientated to inhibit important cytokines as key regulators of the inflammatory process by using different promising heteroaromatic compounds such as quinoxalines, which are cheap and easily synthesized.
420
www.elsevier.com/locate/nbt
During the last years oils and fats are being considered as renewable raw materials for the chemical industry. Therefore, they are a promising alternative to the undesired, limited and pollutants fossil oils. Among these fats and oils, microbial oils are gaining importance due to their advantages over animal fats, vegetable oils and algae oils. Just to cite some examples, they are not affected by climate and season conditions, they can use industrial waste products as inexpensive carbon sources and they can be engineered to be enriched in specific oils that can be directly used as lubricants, functional polymers and other high-value fine chemicals, such as pharmaceuticals and nutraceuticals [1]. Our research is focus in the use of systems metabolic engineering techniques (systems biology, synthetic biology and mathematical models [2]) in the industrial fungus Ashbya gossypii. Firstly, we constructed a strain able to accumulate up to 70% of its cell dry weight as lipids, three times more than the wild type, through the heterologous expression of genes from lipogenic organisms [3]. Secondly, we generated a wide range of engineered strains able to accumulate different fatty acids, which can be used for different biotechnological purposes such as the production of omega 3 fatty acids, waxes, lubricants and biodiesel [4]. We can therefore conclude that A. gossypii is a novel and promising tool for bio-oils production. References [1]. Ledesma-Amaro. Eur J Lipid Sci Technol 2015;117(2):141–4. [2]. Ledesma-Amaro R, Kerkhoven EJ, Revuelta JL, Nielsen J. Biotechnol Bioeng 2014;111(6):1191–9.
POSTERS
PS – R11
2. New therapeutic and diagnostic strategies A role for keratinocytes in developmental myelopoiesis Francisco Juan Martínez-Navarro ∗ , Diana García-Moreno, Azucena López Mu˜noz, José Meseguer, Victoriano Mulero Departamento de Biología Celular e Histología, Facultad de Biología, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
E-mail address: [email protected] (F.J. Martínez-Navarro). Tnfa is a powerful pro-inflammatory cytokine which signals through two receptors, namely Tnfr1 and Tnfr2. Tnfr2 has been shown to protect skin from oxidative stress-induced inflammation using the zebrafish as an experimental model. In the present study, using fluorescent reporter transgene, fluorescence microscopy and RT-qPCR, we studied the impact of Tnfr2 overexpression in zebrafish keratinocytes. Surprisingly, overexpression of Tnfr2 in keratinocytes led to increase neutrophil numbers in 3 days postfertilization larvae concomitantly with decreased transcript levels of Tnfa and increased levels of Il1b, Pu.1, Gata1, Mcsf and Gcsf. These results suggest a critical importance of keratinocytes in developmental myelopoiesis and may explain the systemic neutrophilia associated to several skin inflammatory disorders, such as the acute febrile neutrophilic dermatosis or Sweet’s syndrome.
Following this strategy, we have previously reported how a new collection of 4-alkoxy-6,9-dichloro[1,2,4]triazolo[4,3a]quinoxalines exhibit potent in vitro anti-inflammatory activity as inhibitors of the pro-inflammatory cytokines IL-6 and TNF-␣. Our main aim now is to study how these quinoxaline derivatives act upon different intracellular signaling mechanisms, in an attempt to find their potential molecular targets. We have identified a group of intracellular molecules whose patterns of activation and/or expression are altered by the analyzed compounds, providing further information about the inhibition of TNF-␣ and IL-6. Our results may be of great interest for the pharmaceutical industry, as a starting point for the development of new and more potent anti-inflammatory drugs derived from quinoxalines. http://dx.doi.org/10.1016/j.nbt.2015.10.018
PS – R13
3. Bioenergy and bioremediation Synthetic biology and metabolic engineering to produce microbial oils: towards a sustainable source of fuels, chemicals and nutraceuticals Rodrigo Ledesma-Amaro 1,∗ , Jose Luis Revuelta 2 1
2
INRA, AgroParisTech, UMR1319 Micalis, F-78350 Jouy-en-Josas, France Universidad de Salamanca, Salamanca, Spain
http://dx.doi.org/10.1016/j.nbt.2015.10.017
PS – R12
2. New therapeutic and diagnostic strategies Discovering the molecular targets of 4-alkoxy-6,9dichloro[1,2,4]triazolo[4,3-a]quinoxalines that mediate an anti-inflammatory effect on the macrophage function María Tristán-Manzano 1,∗ , Antonio Guirado 2 , María MartínezEsparza 1 , Pilar García-Pe˜narrubia 1 , Antonio J. Ruiz-Alcaraz 1 1
Department of Biochemistry, Molecular Biology and Immunology B, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain 2 Department of Organic Chemistry, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain
E-mail address: [email protected] (M. Tristán-Manzano). A dysregulation of the inflammatory process, which is a complex biological response directed by the immune system to maintain homeostasis, leads to the development of chronic inflammatory or autoimmune diseases, whose current treatments present many side effects. Nowadays, the search of new therapeutic alternatives focuses on different strategies. Some of them are orientated to inhibit important cytokines as key regulators of the inflammatory process by using different promising heteroaromatic compounds such as quinoxalines, which are cheap and easily synthesized.
420
www.elsevier.com/locate/nbt
During the last years oils and fats are being considered as renewable raw materials for the chemical industry. Therefore, they are a promising alternative to the undesired, limited and pollutants fossil oils. Among these fats and oils, microbial oils are gaining importance due to their advantages over animal fats, vegetable oils and algae oils. Just to cite some examples, they are not affected by climate and season conditions, they can use industrial waste products as inexpensive carbon sources and they can be engineered to be enriched in specific oils that can be directly used as lubricants, functional polymers and other high-value fine chemicals, such as pharmaceuticals and nutraceuticals [1]. Our research is focus in the use of systems metabolic engineering techniques (systems biology, synthetic biology and mathematical models [2]) in the industrial fungus Ashbya gossypii. Firstly, we constructed a strain able to accumulate up to 70% of its cell dry weight as lipids, three times more than the wild type, through the heterologous expression of genes from lipogenic organisms [3]. Secondly, we generated a wide range of engineered strains able to accumulate different fatty acids, which can be used for different biotechnological purposes such as the production of omega 3 fatty acids, waxes, lubricants and biodiesel [4]. We can therefore conclude that A. gossypii is a novel and promising tool for bio-oils production. References [1]. Ledesma-Amaro. Eur J Lipid Sci Technol 2015;117(2):141–4. [2]. Ledesma-Amaro R, Kerkhoven EJ, Revuelta JL, Nielsen J. Biotechnol Bioeng 2014;111(6):1191–9.