Final Report Summary - MICAG (Microbial-induced angiogenesis in the gut)
Summary description of the project objectives:
The mammalian gut is colonised by trillions of bacteria. These gut microbes (microbiota) have coevolved with the host and developed traits that affects host physiology. Germ-free (GF) mice are powerful tools to resolve the effect of the commensal microbiota on the host organism. GF mice are raised under completely sterile conditions and therefore completely lack a gut microbiota. Interestingly, GF mice exhibit arrested intestinal vascularisation if compared to age-matched littermates that were raised in a non-sterile environment (conventional-raised (CONV-R)). Intestinal colonisation with a normal microbiota initiates massive formation of capillary networks in the intestine of ex-GF mice (conventional-derived (CONV-D)). This finding implies that colonisation of the gut with microbial communities evokes the expression of a pro-angiogenic gene repertoire in the intestinal mucosa. The underlying molecular mechanisms that lead to formation of intricate capillary networks are completely unknown and therefore intense research is needed to elucidate how gut microbes affect intestinal morphology and trigger the formation of capillary networks. In the project 'Microbial-induced angiogenesis in the gut' we have dissected pro-angiogenic molecules that can be induced by colonisation with a gut microbiota and thus regulate microvessel formation in the intestinal mucosa. A detailed understanding on how microbes affect intestinal morphogenesis and in particular the formation of microvessels in the mucosa is of major interest in clinical medicine, since a disturbed formation of microvessels is observed in inflammatory bowel disease (IBD) - a disease with unknown etiology. In summary, all objectives of the project were achieved.
Description of the work carried out
After reintegration at the host laboratory at the University Clinics Würzburg and starting the project at the Institute of Clinical Biochemistry and Pathobiochemistry in Würzburg / Germany intestinal tissues from GF, CONV-R and CONV-D mice were shipped from the Laboratroy of Prof. Fredrik Bäckhed to Würzburg / Germany. The described expression analyses for murine CD31 and the expression of various angiogenic factors (vascular endothelial growth factor (VEGF) system, Tie-2 / Ang-1 pathway) in the small intestinal mucosa were performed by quantitative RT-PCR. We also analysed immunofluorescence stainings of the vascular marker CD31 in frozen sections of the small intestine by immunohistochemistry. Since the angiogenic factor angiopoietin-1 (Ang-1) that is required for vessel maturation was found to be regulated by the extrinsic coagulation pathway (tissue factor pathway) we performed Western blot analyses and functional analyses on the coagulation initiator protein tissue factor. Indeed we found differences in procoagulant activity comparing GF and colonised small intestinal tissue lysates. Since coagulation factors possess a known signalling function via protease-activated receptors (PARs), we reasoned that differences in coagulation could be the cause of the increased vascularisation in the small intestinal villus structures of CONV-R and CONV-D mice and thus we extended our analyses and isolated primary enterocytes from GF and CONV-R mice. Upon microbial colonisation, we found altered PAR-1 expression levels in the enterocytes. For this reason, we initiated an ongoing collaboration with Prof. Wolfram Ruf (Scripps Research Institute, La Jolla, United States (US)), who is amongst the leading experts in the area of coagulation factor signalling. For our project, he provided us with small intestinal tissues from gene targeted PAR-1 and PAR-2 deficient mice and the respective wild-type (WT) controls. We analysed tissue specimens from these mice for differences in vascularisation (both by immunohistochemistry and qualitative real-time polymerase chain reaction (qRT-PCR)) and the angiogenic factor Ang-1. This work resulted in a first author publication in the top-tier journal Nature (Reinhardt C et al., Tissue factor and PAR1promote microbiota-induced intestinal vascular remodelling. Nature 2012; 483:627-31).
Description of main results achieved
The planned immunohistochemical analysis of the vascular marker protein PECAM-1 (CD31) in small intestine tissues has been successfully carried out and a profound difference in the vascularisation of GF, CONV-D and CONV-R mice was found by image analysis of villus capillaries. This has also been confirmed by qRT-PCR analysis with primers that are specific for murine CD31. These experiments have been described as Aim 1 in the original proposal. We also succeeded in identifying certain proangiogenic factors that are regulated by colonisation with a gut microbiota. Importantly, expression of Ang-1 and phosphorylation of its tyrosine kinase receptor Tie-2 that is involved in vessel maturation was altered by colonisation with a gut microbiota (Reinhardt C et al., Nature, 2012). Inhibition of this angiogenic signalling pathway in colonised mice with the peptibody mL4-3 resulted in decreased formation of capillary networks in the villus structures. Hence, we could show that microbial regulation of the Ang-1 / Tie-2 signalling pathway is of functional importance. Amongst the proangiogenic factors analysed, we found differences in the tissue factor protein between GF, CONV-D and CONV-R mice. The differences observed might be due to posttranslational protein modification. Moreover, expression levels of the thrombin receptor PAR-1 was significantly elevated in CONV-R mice. However, messenger ribonucleic acid (mRNA) messages of other factors like VEGF-A, VEGFR-2, CXCL-12, PDGF-BB, and Tie-2 remained unaffected by microbial colonisation suggesting that the VEGF pathway is of less importance in mucosal vascular remodelling. Further analyses on the expression of other proangiogenic factors found regulated by colonisation with a gut microbiota and on downstream kinase pathways are in progress. This resulted in the identification of additional target molecules that are regulated by the gut microbiota. Thus, we were able to achieve all major goals that are mentioned under Aim 2 in the original proposal.
At present, further immunohistochemistry experiments are done by the Doctor of Philosophy (PhD) student Klytaimnistra Kiouptsi, a biotechnologist who was recruited to the project from the University of Ioannina in Greece. The analysis of signalling complexes were successfully completed and we could demonstrate an interaction of tissue factor with integrin beta 1 in the small intestinal epithelium using immunoprecipitation and the DuoLink in situ proximity ligation assay to study the interaction of tissue factor and the integrin ß1 chain (Reinhardt C et al., Nature, 2012). These experiments are summarised in aim 3 of the original grant proposal and we could perform all experiments and also fulfil aim 3 in the given timeline.
Conclusions
During the first year of the project, at the Institute of Clinical Biochemistry and Pathobiochemistry in Würzburg, we identified a novel microbiota-induced pathway that links coagulation factor signalling to the Ang-1 / Tie-2 signalling axis and might thus explain how the extrinsic coagulation pathway contributes to vascular remodelling in the small intestinal mucosa. We further continued this project after the junior research group has moved to the Centre for Thrombosis and Hemostasis (CTH) in Mainz in July 2011 when the Marie Curie Reintegration Grant (ERG) was transferred from Würzburg to Mainz. The results of this project were included into a manuscript that has recently been published in the top-tier journal Nature (Reinhardt C et al., Nature, 2012; Impact Factor: 36.28). Since increased angiogenesis is described as a hallmark in the inflamed mucosa in Crohn's disease these mechanisms could play a role during pathogenesis. Moreover, the maturation of capillaries in small intestinal villus structures via coagulation factor signalling might occur upon colonisation of infants after birth. Now, that we have gained profound mechanistic insights on how the gut microbiota regulates formation of capillary networks we will analyse intestinal angiogenesis during colitis in future projects that could be initiated by this ERG. We are about to successfully finish the Marie ERG project within the envisaged timescale of 3 years and we could achieve all aims defined in the original grant proposal.
Socio-economic impacts of the project
PhD projects and career development
With this European Union (EU)-funded project the PhD project of Ms Nives Hörmann within the junior research group of Christoph Reinhardt could initially be financed. Nives is a very skilled molecular biotechnologist who was recruited from the RWTH Aachen to Würzburg. To gain insights into the rapidly evolving field of host-microbial interactions she visited the FEMS Congress 2011 in Geneva / Switzerland where she could make the acquaintance of Prof. Bäckhed, a collaborator within the project and the former mentor of Dr Reinhardt. She also has moved to Mainz to continue this interesting line of research within the junior group of C. Reinhardt where her position will continuously be funded from other sources. For the required material costs we could successfully apply for a project contribution by the Universitätsbund Würzburg e.V. (EUR 4800). Moreover, this grant enabled Nives Hörmann to successfully take part in a course in experimental animal science according to FELASA B that was held in Würzburg in July 2011. Nives Hörmann had the opportunity to present her results in form of a poster at the International Vascular Biology Meeting 2012 in Wiesbaden and in form of an oral presentation at the Annual Meeting of the Society for Thrombosis and Hemostasis Research taking place at 20 - 23 February in Munich, Germany. Furthermore, an additional PhD student could be recruited to the junior research group of Dr Reinhardt in May 2012 when the remaining grant sum was approved for transfer from Würzburg to Mainz. Klytaimnistra Kiouptsi, a skilled Biotechnologist from the University of Ioannina in Greece was employed within the project and she could perform immunohistochemistry analyses that are the basis of further grant applications for additional third party funding (e.g. German Research Council, DFG). From January 2013, the employment of Klytaimnistra Kiouptsi in the junior research group was continued with other intramural project grants and preliminary data of the ERG were essential for successful grant application. Both Nives Hörmann and Klytaimnistra Kiouptsi were accepted as doctoral candidate fellows at the Centre for Thrombosis and Hemostasis and thus a sound PhD education with various clinical interactions is assured (see: http://www.cth-mainz.de/cth/career-support/doctoral-candidate-program.html?L=1 for details).
The EU-funded project enabled Dr Christoph Reinhardt to develop an independent line of research that is essential to create an own research programme as an independent junior group leader (see CTH website: http://www.cth-mainz.de/cth/professorships-research-groups/juniorgroup-reinhardt-exp-research.html?L=1 online for further details). The results acquired during the first year of the project strengthened the competitiveness of Dr Reinhardt to obtain a junior research position at the Centre for Thrombosis and Hemostasis (CTH) in Mainz / Germany. The CTH Mainz is one of eight health care centres that were funded by the German Federal Ministry for Education and Research (BMBF) until August 2015 and a further five year prolongation of this centre grant is envisaged. This ensured a long-term job perspective for the PhD student Nives Hörmann (until November 2013) and Dr Reinhardt (until August 2015).
Importantly, the ERG project led to successful applications of third party funding (1 PhD position, EUR 112 000) from the foundation 'Pathobiochemistry and Molecular Diagnostics' (Deutsche Vereinte Gesellschaft für Klinische Chemie und Laboratoriumsmedizin) and from the German Research Council, DFG (1 post-doc position, EUR 188 000). This funding of innovative projects is critical to consolidate the junior research group of Dr Christoph Reinhardt.
The project's results could be presented by an oral presentation of Dr Reinhardt at the International Vascular Biology Meeting (IVBM) 2012 in Wiesbaden / Germany. Furthermore, Dr Reinhardt's published work has been awarded with the prestigious Hermann-Rein-Award on behalf of the Gesellschaft für Mikrozirkulation und Vaskuläre Biologie to Dr. Christoph Reinhardt (see http://www.gfmvb.com/preistrager.htm online for further details). On the basis of his scientific work, Dr Reinhardt is presently considered for a W2 professorship for 'Vascular Immunotherapy' in Munich (Listenplatz at the Institut für Prophylaxe und Epidemiologie der Kreislauferkrankungen) and a W1 Professorship in 'Membrane Biochemistry' in Mainz (Listenplatz at the Institut für Biochemie) where he was invited to present his work. This clearly demonstrates that the EU ERG has strongly boosted the career path of Dr C. Reinhardt.
In Mainz, Dr Reinhardt and his group have successfully established own isolator systems for the breeding and maintenance of germfree mouse colonies and this is a further example how applicable knowledge has been transferred in the continued collaboration with Dr Reinhardt's former mentor Prof. Fredrik Bäckhed (Gothenburg, Sweden). The establishment of mouse isolator technology has been strongly supported by Prof. Kurt Reifenberg (former head of the Central Animal Experimentation Facility / ZVTE, University of Mainz). Dr Reinhardt and his team recently succeeded in first re-derivations of transgene mice as germfree which shows that the facility is fully operational.
Networking with other scientists
With this project that is mainly based on germfree mouse technology a long-term collaboration with Prof. Fredrik Bäckhed in Gothenburg / Sweden was established. The PhD student employed within the project met with Prof. Bäckhed at the FEMS conference in Geneva / Switzerland and he invited Dr Christoph Reinhardt and Nives Hörmann to visit his germfree mouse facility in Gothenburg to experience an intense training required to establish own isolators with germfree mice in Mainz / Germany. Prof. Bäckhed also visited the CTH Mainz in August 2012 and presented work from his group in Gothenburg. Furthermore, from his time as a Marie Curie post-doctoral fellow his research team has ongoing collaborations with Prof. Agnes Wold (Institute for Clinical Bacteriology, Gothenburg, Sweden) regarding the monocolonisation of germfree mice with isogenic Staphylococcus aureus mutants in the germfree mouse facility in Mainz.
Moreover, Dr Reinhardt has ongoing collaborations with Prof. Wolfram Ruf from the Scripps Institute in La Jolla / US Prof. Ruf is one of the pioneers in the tissue factor field. He provided us with antibody reagents and snap frozen small intestinal tissues from gene targeted PAR-deficient mice and WT controls. He will join the newly established Center for Thrombosis and Hemostais (CTH) in Mainz from 2014 as a well-endowed Humboldt-Professor and this will certainly support the ongoing work and the research field of Dr. Reinhardt on the role of the microbiota on the clotting system.
In Würzburg, Dr Reinhardt shared his office with Dr Erik Henke, an experienced researcher in the angiogenesis field who moved from the Sloan Kettering Cancer Centre in New York to Würzburg. During the first couple of months in Würzburg, Dr Reinhardt could also make the acquaintance of Prof. Bernhard Nieswandt (Director of the Virchow Research Center), Prof. Jens Waschke (Institute for Anatomy, now in Munich) and of Dr Alma Zernecke (now in Munich), all leading scientists in the Vascular Biology field.
In Mainz, Dr Reinhardt has ongoing collaborations with Dr Philip Wenzel (Cardiology), Prof. Kerstin Steinbrink (Dermatology Clinics) and Dr Natalia Soshnikova (Institute of Molecular Biology, Boehringer Ingelheim) who are highly interested in the germfree mouse technology that Dr Reinhardt and his team have established. Furthermore, Dr Reinhardt has agreed to support the group of Prof. John Baines at the May-Planck-Institute for Evolutionary Biology to set up own mouse isolator systems in Plön / Germany. The group of Dr Reinhardt has further interactions with Prof. Andres Hidalgo from the Centro Nacional de Investigationes Cardiovasculares (Madrid, Spain), Prof. Deborah Gumucio (University of Michigan, US) and with Daniela Wenzel (Institute of Physiology I, Bonn, Germany).
The mammalian gut is colonised by trillions of bacteria. These gut microbes (microbiota) have coevolved with the host and developed traits that affects host physiology. Germ-free (GF) mice are powerful tools to resolve the effect of the commensal microbiota on the host organism. GF mice are raised under completely sterile conditions and therefore completely lack a gut microbiota. Interestingly, GF mice exhibit arrested intestinal vascularisation if compared to age-matched littermates that were raised in a non-sterile environment (conventional-raised (CONV-R)). Intestinal colonisation with a normal microbiota initiates massive formation of capillary networks in the intestine of ex-GF mice (conventional-derived (CONV-D)). This finding implies that colonisation of the gut with microbial communities evokes the expression of a pro-angiogenic gene repertoire in the intestinal mucosa. The underlying molecular mechanisms that lead to formation of intricate capillary networks are completely unknown and therefore intense research is needed to elucidate how gut microbes affect intestinal morphology and trigger the formation of capillary networks. In the project 'Microbial-induced angiogenesis in the gut' we have dissected pro-angiogenic molecules that can be induced by colonisation with a gut microbiota and thus regulate microvessel formation in the intestinal mucosa. A detailed understanding on how microbes affect intestinal morphogenesis and in particular the formation of microvessels in the mucosa is of major interest in clinical medicine, since a disturbed formation of microvessels is observed in inflammatory bowel disease (IBD) - a disease with unknown etiology. In summary, all objectives of the project were achieved.
Description of the work carried out
After reintegration at the host laboratory at the University Clinics Würzburg and starting the project at the Institute of Clinical Biochemistry and Pathobiochemistry in Würzburg / Germany intestinal tissues from GF, CONV-R and CONV-D mice were shipped from the Laboratroy of Prof. Fredrik Bäckhed to Würzburg / Germany. The described expression analyses for murine CD31 and the expression of various angiogenic factors (vascular endothelial growth factor (VEGF) system, Tie-2 / Ang-1 pathway) in the small intestinal mucosa were performed by quantitative RT-PCR. We also analysed immunofluorescence stainings of the vascular marker CD31 in frozen sections of the small intestine by immunohistochemistry. Since the angiogenic factor angiopoietin-1 (Ang-1) that is required for vessel maturation was found to be regulated by the extrinsic coagulation pathway (tissue factor pathway) we performed Western blot analyses and functional analyses on the coagulation initiator protein tissue factor. Indeed we found differences in procoagulant activity comparing GF and colonised small intestinal tissue lysates. Since coagulation factors possess a known signalling function via protease-activated receptors (PARs), we reasoned that differences in coagulation could be the cause of the increased vascularisation in the small intestinal villus structures of CONV-R and CONV-D mice and thus we extended our analyses and isolated primary enterocytes from GF and CONV-R mice. Upon microbial colonisation, we found altered PAR-1 expression levels in the enterocytes. For this reason, we initiated an ongoing collaboration with Prof. Wolfram Ruf (Scripps Research Institute, La Jolla, United States (US)), who is amongst the leading experts in the area of coagulation factor signalling. For our project, he provided us with small intestinal tissues from gene targeted PAR-1 and PAR-2 deficient mice and the respective wild-type (WT) controls. We analysed tissue specimens from these mice for differences in vascularisation (both by immunohistochemistry and qualitative real-time polymerase chain reaction (qRT-PCR)) and the angiogenic factor Ang-1. This work resulted in a first author publication in the top-tier journal Nature (Reinhardt C et al., Tissue factor and PAR1promote microbiota-induced intestinal vascular remodelling. Nature 2012; 483:627-31).
Description of main results achieved
The planned immunohistochemical analysis of the vascular marker protein PECAM-1 (CD31) in small intestine tissues has been successfully carried out and a profound difference in the vascularisation of GF, CONV-D and CONV-R mice was found by image analysis of villus capillaries. This has also been confirmed by qRT-PCR analysis with primers that are specific for murine CD31. These experiments have been described as Aim 1 in the original proposal. We also succeeded in identifying certain proangiogenic factors that are regulated by colonisation with a gut microbiota. Importantly, expression of Ang-1 and phosphorylation of its tyrosine kinase receptor Tie-2 that is involved in vessel maturation was altered by colonisation with a gut microbiota (Reinhardt C et al., Nature, 2012). Inhibition of this angiogenic signalling pathway in colonised mice with the peptibody mL4-3 resulted in decreased formation of capillary networks in the villus structures. Hence, we could show that microbial regulation of the Ang-1 / Tie-2 signalling pathway is of functional importance. Amongst the proangiogenic factors analysed, we found differences in the tissue factor protein between GF, CONV-D and CONV-R mice. The differences observed might be due to posttranslational protein modification. Moreover, expression levels of the thrombin receptor PAR-1 was significantly elevated in CONV-R mice. However, messenger ribonucleic acid (mRNA) messages of other factors like VEGF-A, VEGFR-2, CXCL-12, PDGF-BB, and Tie-2 remained unaffected by microbial colonisation suggesting that the VEGF pathway is of less importance in mucosal vascular remodelling. Further analyses on the expression of other proangiogenic factors found regulated by colonisation with a gut microbiota and on downstream kinase pathways are in progress. This resulted in the identification of additional target molecules that are regulated by the gut microbiota. Thus, we were able to achieve all major goals that are mentioned under Aim 2 in the original proposal.
At present, further immunohistochemistry experiments are done by the Doctor of Philosophy (PhD) student Klytaimnistra Kiouptsi, a biotechnologist who was recruited to the project from the University of Ioannina in Greece. The analysis of signalling complexes were successfully completed and we could demonstrate an interaction of tissue factor with integrin beta 1 in the small intestinal epithelium using immunoprecipitation and the DuoLink in situ proximity ligation assay to study the interaction of tissue factor and the integrin ß1 chain (Reinhardt C et al., Nature, 2012). These experiments are summarised in aim 3 of the original grant proposal and we could perform all experiments and also fulfil aim 3 in the given timeline.
Conclusions
During the first year of the project, at the Institute of Clinical Biochemistry and Pathobiochemistry in Würzburg, we identified a novel microbiota-induced pathway that links coagulation factor signalling to the Ang-1 / Tie-2 signalling axis and might thus explain how the extrinsic coagulation pathway contributes to vascular remodelling in the small intestinal mucosa. We further continued this project after the junior research group has moved to the Centre for Thrombosis and Hemostasis (CTH) in Mainz in July 2011 when the Marie Curie Reintegration Grant (ERG) was transferred from Würzburg to Mainz. The results of this project were included into a manuscript that has recently been published in the top-tier journal Nature (Reinhardt C et al., Nature, 2012; Impact Factor: 36.28). Since increased angiogenesis is described as a hallmark in the inflamed mucosa in Crohn's disease these mechanisms could play a role during pathogenesis. Moreover, the maturation of capillaries in small intestinal villus structures via coagulation factor signalling might occur upon colonisation of infants after birth. Now, that we have gained profound mechanistic insights on how the gut microbiota regulates formation of capillary networks we will analyse intestinal angiogenesis during colitis in future projects that could be initiated by this ERG. We are about to successfully finish the Marie ERG project within the envisaged timescale of 3 years and we could achieve all aims defined in the original grant proposal.
Socio-economic impacts of the project
PhD projects and career development
With this European Union (EU)-funded project the PhD project of Ms Nives Hörmann within the junior research group of Christoph Reinhardt could initially be financed. Nives is a very skilled molecular biotechnologist who was recruited from the RWTH Aachen to Würzburg. To gain insights into the rapidly evolving field of host-microbial interactions she visited the FEMS Congress 2011 in Geneva / Switzerland where she could make the acquaintance of Prof. Bäckhed, a collaborator within the project and the former mentor of Dr Reinhardt. She also has moved to Mainz to continue this interesting line of research within the junior group of C. Reinhardt where her position will continuously be funded from other sources. For the required material costs we could successfully apply for a project contribution by the Universitätsbund Würzburg e.V. (EUR 4800). Moreover, this grant enabled Nives Hörmann to successfully take part in a course in experimental animal science according to FELASA B that was held in Würzburg in July 2011. Nives Hörmann had the opportunity to present her results in form of a poster at the International Vascular Biology Meeting 2012 in Wiesbaden and in form of an oral presentation at the Annual Meeting of the Society for Thrombosis and Hemostasis Research taking place at 20 - 23 February in Munich, Germany. Furthermore, an additional PhD student could be recruited to the junior research group of Dr Reinhardt in May 2012 when the remaining grant sum was approved for transfer from Würzburg to Mainz. Klytaimnistra Kiouptsi, a skilled Biotechnologist from the University of Ioannina in Greece was employed within the project and she could perform immunohistochemistry analyses that are the basis of further grant applications for additional third party funding (e.g. German Research Council, DFG). From January 2013, the employment of Klytaimnistra Kiouptsi in the junior research group was continued with other intramural project grants and preliminary data of the ERG were essential for successful grant application. Both Nives Hörmann and Klytaimnistra Kiouptsi were accepted as doctoral candidate fellows at the Centre for Thrombosis and Hemostasis and thus a sound PhD education with various clinical interactions is assured (see: http://www.cth-mainz.de/cth/career-support/doctoral-candidate-program.html?L=1 for details).
The EU-funded project enabled Dr Christoph Reinhardt to develop an independent line of research that is essential to create an own research programme as an independent junior group leader (see CTH website: http://www.cth-mainz.de/cth/professorships-research-groups/juniorgroup-reinhardt-exp-research.html?L=1 online for further details). The results acquired during the first year of the project strengthened the competitiveness of Dr Reinhardt to obtain a junior research position at the Centre for Thrombosis and Hemostasis (CTH) in Mainz / Germany. The CTH Mainz is one of eight health care centres that were funded by the German Federal Ministry for Education and Research (BMBF) until August 2015 and a further five year prolongation of this centre grant is envisaged. This ensured a long-term job perspective for the PhD student Nives Hörmann (until November 2013) and Dr Reinhardt (until August 2015).
Importantly, the ERG project led to successful applications of third party funding (1 PhD position, EUR 112 000) from the foundation 'Pathobiochemistry and Molecular Diagnostics' (Deutsche Vereinte Gesellschaft für Klinische Chemie und Laboratoriumsmedizin) and from the German Research Council, DFG (1 post-doc position, EUR 188 000). This funding of innovative projects is critical to consolidate the junior research group of Dr Christoph Reinhardt.
The project's results could be presented by an oral presentation of Dr Reinhardt at the International Vascular Biology Meeting (IVBM) 2012 in Wiesbaden / Germany. Furthermore, Dr Reinhardt's published work has been awarded with the prestigious Hermann-Rein-Award on behalf of the Gesellschaft für Mikrozirkulation und Vaskuläre Biologie to Dr. Christoph Reinhardt (see http://www.gfmvb.com/preistrager.htm online for further details). On the basis of his scientific work, Dr Reinhardt is presently considered for a W2 professorship for 'Vascular Immunotherapy' in Munich (Listenplatz at the Institut für Prophylaxe und Epidemiologie der Kreislauferkrankungen) and a W1 Professorship in 'Membrane Biochemistry' in Mainz (Listenplatz at the Institut für Biochemie) where he was invited to present his work. This clearly demonstrates that the EU ERG has strongly boosted the career path of Dr C. Reinhardt.
In Mainz, Dr Reinhardt and his group have successfully established own isolator systems for the breeding and maintenance of germfree mouse colonies and this is a further example how applicable knowledge has been transferred in the continued collaboration with Dr Reinhardt's former mentor Prof. Fredrik Bäckhed (Gothenburg, Sweden). The establishment of mouse isolator technology has been strongly supported by Prof. Kurt Reifenberg (former head of the Central Animal Experimentation Facility / ZVTE, University of Mainz). Dr Reinhardt and his team recently succeeded in first re-derivations of transgene mice as germfree which shows that the facility is fully operational.
Networking with other scientists
With this project that is mainly based on germfree mouse technology a long-term collaboration with Prof. Fredrik Bäckhed in Gothenburg / Sweden was established. The PhD student employed within the project met with Prof. Bäckhed at the FEMS conference in Geneva / Switzerland and he invited Dr Christoph Reinhardt and Nives Hörmann to visit his germfree mouse facility in Gothenburg to experience an intense training required to establish own isolators with germfree mice in Mainz / Germany. Prof. Bäckhed also visited the CTH Mainz in August 2012 and presented work from his group in Gothenburg. Furthermore, from his time as a Marie Curie post-doctoral fellow his research team has ongoing collaborations with Prof. Agnes Wold (Institute for Clinical Bacteriology, Gothenburg, Sweden) regarding the monocolonisation of germfree mice with isogenic Staphylococcus aureus mutants in the germfree mouse facility in Mainz.
Moreover, Dr Reinhardt has ongoing collaborations with Prof. Wolfram Ruf from the Scripps Institute in La Jolla / US Prof. Ruf is one of the pioneers in the tissue factor field. He provided us with antibody reagents and snap frozen small intestinal tissues from gene targeted PAR-deficient mice and WT controls. He will join the newly established Center for Thrombosis and Hemostais (CTH) in Mainz from 2014 as a well-endowed Humboldt-Professor and this will certainly support the ongoing work and the research field of Dr. Reinhardt on the role of the microbiota on the clotting system.
In Würzburg, Dr Reinhardt shared his office with Dr Erik Henke, an experienced researcher in the angiogenesis field who moved from the Sloan Kettering Cancer Centre in New York to Würzburg. During the first couple of months in Würzburg, Dr Reinhardt could also make the acquaintance of Prof. Bernhard Nieswandt (Director of the Virchow Research Center), Prof. Jens Waschke (Institute for Anatomy, now in Munich) and of Dr Alma Zernecke (now in Munich), all leading scientists in the Vascular Biology field.
In Mainz, Dr Reinhardt has ongoing collaborations with Dr Philip Wenzel (Cardiology), Prof. Kerstin Steinbrink (Dermatology Clinics) and Dr Natalia Soshnikova (Institute of Molecular Biology, Boehringer Ingelheim) who are highly interested in the germfree mouse technology that Dr Reinhardt and his team have established. Furthermore, Dr Reinhardt has agreed to support the group of Prof. John Baines at the May-Planck-Institute for Evolutionary Biology to set up own mouse isolator systems in Plön / Germany. The group of Dr Reinhardt has further interactions with Prof. Andres Hidalgo from the Centro Nacional de Investigationes Cardiovasculares (Madrid, Spain), Prof. Deborah Gumucio (University of Michigan, US) and with Daniela Wenzel (Institute of Physiology I, Bonn, Germany).