Institut für Molekulare Medizin Mainz
AG Mufazalov
Research
We are determined to decipher mechanisms leading to immune dysfunction. In particular, our research focuses on the role of cytokine signaling in autoimmunity, chronic inflammation and maternal immune activation during pregnancy. We are confident that the immune system plays a central role in keeping our physiology in check and the majority of diseases could be treated by restoration of disrupted immune homeostasis. Through our research, we aim to improve existing and develop novel therapeutic strategies To address complex biomedical questions, we utilize preclinical models that are based on genetically modified mice with cell-type-specific gene knockouts and cytokine overexpression. We routinely employ in vivo immunizations, adoptive cell transfers, timed pregnancies, behavioral tests and other protocols. Our standard methods include, but are not limited to, Flow Cytometry, Western Blotting, ELISA, qPCR and primary cell cultures. Our group works in close collaboration with other members of the Institute for Molecular Medicine, especially with the AG Waisman.
Interleukin-1 (IL-1) signaling is strongly implicated in numerous pathologies, including central nervous system (CNS) disorders such as multiple sclerosis (MS), Alzheimer's disease and Parkinson's disease. Hallmarks of MS are disseminated lesions caused by immune cell infiltration into the CNS which promotes inflammation, demyelination, gliosis and neuroaxonal degeneration. In severe cases, MS patients suffer from sensory/visual disturbances, motor impairments, fatigue, pain and cognitive deficits. Elevated levels of IL-1 have been reported within the white matter and in acute lesions of MS patients. Typically, increased IL-1 activity is associated with more severe MS symptoms. In this regard, therapeutic targeting of the IL-1 pathway is an attractive solution to pursue, which, however, requires additional experimental support.
To mimic Multiple Sclerosis in a preclinical setting, we use a murine model for CNS-directed autoimmunity, namely Experimental Autoimmune Encephalomyelitis (EAE).
The assessment of disease progression and analysis of CNS infiltrates in interleukin-1 receptor type 1 (IL-1R1) as well as interleukin-1 receptor type 2 (IL-1R2) deficient mice is our toolbox in studying the role of IL-1 in CNS-directed autoimmunity.
To mimic Multiple Sclerosis in a preclinical setting, we use a murine model for CNS-directed autoimmunity, namely Experimental Autoimmune Encephalomyelitis (EAE).
The assessment of disease progression and analysis of CNS infiltrates in interleukin-1 receptor type 1 (IL-1R1) as well as interleukin-1 receptor type 2 (IL-1R2) deficient mice is our toolbox in studying the role of IL-1 in CNS-directed autoimmunity.
2) The role of interleukin-6 signaling in the development of lymphoproliferative disorders (This project is relevant to Idiopathic Multicentric Castleman Disease, a lymphoproliferative disorder with very limited treatment options and with a significant proportion of patient refractory to anti-IL-6 therapy)
Interleukin-6 (IL-6) is a critical cytokine for host immune defense and its elevated levels are found in literally all inflammatory conditions. In extreme cases, such as sepsis or cytokine release syndrome (CRS) caused by SARS-CoV-2 infection or as a side effect of CAR T cell therapy, the excessive levels of IL-6 can cause life-threatening complications.
Idiopathic Multicentric Castleman Disease (iMCD) is a rare, but devastating, lymphoproliferative disorder. In this disease, IL-6 is considered to be a major pathogenic factor that causes lymphadenopathy, hepatosplenomegaly, thrombocytopenia, plasmacytosis and multiorgan dysfunction. First-line symptomatic treatment largely relies on inhibition of IL-6 signaling, however, up to 70% of patients remain refractory to IL-6 blockade. The progress on the development of alternative therapeutic interventions remains limited due to the absence of suitable preclinical models.
In this project we aim to identify novel molecular targets in iMCD and repurpose drugs, which are already approved for other inflammatory disorders. We are also fascinated by the idea of developing gene-edited adoptive cell transfer therapy for the treatment of iMCD. For that, we use mice with conditional IL-6 overexpression that display key features of iMCD and mice deficient in IL-6 signaling.
Idiopathic Multicentric Castleman Disease (iMCD) is a rare, but devastating, lymphoproliferative disorder. In this disease, IL-6 is considered to be a major pathogenic factor that causes lymphadenopathy, hepatosplenomegaly, thrombocytopenia, plasmacytosis and multiorgan dysfunction. First-line symptomatic treatment largely relies on inhibition of IL-6 signaling, however, up to 70% of patients remain refractory to IL-6 blockade. The progress on the development of alternative therapeutic interventions remains limited due to the absence of suitable preclinical models.
In this project we aim to identify novel molecular targets in iMCD and repurpose drugs, which are already approved for other inflammatory disorders. We are also fascinated by the idea of developing gene-edited adoptive cell transfer therapy for the treatment of iMCD. For that, we use mice with conditional IL-6 overexpression that display key features of iMCD and mice deficient in IL-6 signaling.
3) Fitness of Foxp3+ regulatory T (Treg) cells under inflammatory conditions (This project is relevant to a broad range of inflammatory conditions and aims at developing Treg cell-based therapies)
Regulatory T (Treg) cells are a specialized subset of immuno-suppressive CD4 T cells that express the lineage-specific transcription factor Foxp3. Treg cells regulate a variety of immune responses in various biological contexts, including inflammation, cancer, metabolism, tissue repair, pregnancy and more. Humans and mice with impaired Treg cell function suffer from severe-to-lethal autoimmune disorders. Although the developmental pathways leading to the generation of Treg cells are well-studied, little is known how inflammatory mediators (and in particular IL-6) impact mature Treg cells function. Filling up this gap of knowledge is a keystone for improving treatment strategies. Such treatments could modulate Treg cells in patients on-site or could be a part of fast-developing strategies of adoptive cell transfer therapies. Thanks to advances in genetic engineering, Treg cells might be isolated, modified, expanded and transferred back into the patients as living drugs.Immune responses occur at a complex in vivo environment; therefore, they cannot be fully recapitulated by in vitro systems. To study short-term and long-lasting effects of IL-6 exposure on Treg cells, we use transgenic mice with IL-6 overexpression and employ Treg cell depletion, Treg cell modulation and Treg cell transfer protocols. We are confident that our translational approach is highly relevant for the development of Treg cell based therapeutic strategies aiming at suppression of Treg cells to promote anti-tumor immunity, or in opposite, stimulation of Treg cells in autoimmune disorders.
4) Prenatal imprinting of the nervous- and immune system by the pro-inflammatory cytokines IL-6 and IL-17A (This project is relevant to the Autism Spectrum Disorder, a group of neurodevelopmental diseases with no standard-of-care protocols)
Recent discoveries highlighted that maternal immune activation (MIA) in response to infections during pregnancy influences the development of fetal nervous and immune system. Dysregulated signaling of maternal IL-6 and IL-17A are key components of MIA and a high-risk factor for the development of autism spectrum disorders (ASD) in offspring. The core ASD symptoms, which vary in severity, are impaired communication and social skills, restricted or repetitive behaviors and intellectual disability.
Currently, there is no definitive standard-of-care neither for already developed ASD nor for the prevention of ASD development. Progress in the field is prevented by the fact that research and therapeutic interventions during human pregnancy are extremely limited for ethical reasons.
To study the etiology of ASD in response to maternally produced cytokines, we use well-defined transgenic mouse models with IL-6 and IL-17A overexpression during pregnancy. This allows us to characterize long-lasting consequences and decipher mechanisms of prenatal imprinting to IL-6 and IL-17A. We believe that insights from our studies will be used to design novel treatment strategies to prevent the development of ASD.
Currently, there is no definitive standard-of-care neither for already developed ASD nor for the prevention of ASD development. Progress in the field is prevented by the fact that research and therapeutic interventions during human pregnancy are extremely limited for ethical reasons.
To study the etiology of ASD in response to maternally produced cytokines, we use well-defined transgenic mouse models with IL-6 and IL-17A overexpression during pregnancy. This allows us to characterize long-lasting consequences and decipher mechanisms of prenatal imprinting to IL-6 and IL-17A. We believe that insights from our studies will be used to design novel treatment strategies to prevent the development of ASD.
Publications
Selected publications:
For a full publication list see: Pubmed
Andruszewski D, Uhlfelder DC, Desiato G, Regen T, Schelmbauer C, Blanfeld M, Scherer L, Radyushkin K, Pozzi D, Waisman A and Mufazalov IA#. Embryo-restricted responses to maternal IL- 17A promote neurodevelopmental disorders in mouse offspring. Molecular Psychiatry 2024, Online ahead of print. doi: 10.1038/s41380-024-02772-6. # - corresponding author
Regen T, Isaac S, Amorim A, Núñez NG, Hauptmann J, Shanmugavadivu A, Klein M, Sankowski R, Mufazalov IA, Yogev N, Huppert J, Wanke F, Witting M, Grill A, Gálvez EJC, Nikolaev A, Blanfeld M, Prinz I, Schmitt-Kopplin P, Strowig T, Reinhardt C, Prinz M, Bopp T, Becher B, Ubeda C and Waisman A. IL-17 controls central nervous system autoimmunity through the intestinal microbiome. Science Immunology 2021, 6(56):eaaz6563.
Lee CJ, Mehdizadeh S, Smith J, Young A, Mufazalov IA, Mowery CT, Daud A and Bluestone AJ. Regulatory T cell control of systemic immunity and immunotherapy response in liver metastasis. Science Immunology 2020, 5(52):eaba0759.
Mufazalov IA#, Andruszewski D, Schelmbauer C, Heink S, Blanfeld M, Masri J, Tang Y, Schuler R, Eich C, Wunderlich TF, Karbach SH, Bluestone JA, Korn T and Waisman A. Cutting Edge: IL-6-driven immune dysregulation is strictly dependent on IL-6R α-chain expression. Journal of Immunology 2020, 204(4): 747-751. # - corresponding author
Dmitrieva-Posocco O, Dzutsev A, Posocco DF, Hou V, Yuan W, Thovarai V, Mufazalov IA, Gunzer M, Shilovskiy IP, Khaitov MR, Trinchieri G, Waisman A and Grivennikov SI. Cell-Type-Specific Responses to Interleukin-1 Control Microbial Invasion and Tumor-Elicited Inflammation in Colorectal Cancer. Immunity 2019, 50(1):166-180.e7
Atretkhany KSN*, Mufazalov IA*, Dunst J, Kuchmiy A, Gogoleva VS, Andruszewski D, Drutskaya MS, Faustman DL, Schwabenland M, Prinz M, Kruglov AA, Waisman A and Nedospasov SA. Intrinsic TNFR2 signaling in T regulatory cells provides protection in CNS autoimmunity. Proc Natl Acad Sci USA 2018, 115(51):13051-13056. *co-first authorship
Schlüter T, Schelmbauer C, Karram K and Mufazalov IA#. Regulation of IL-1 signaling by the decoy receptor IL-1R2. Journal of Molecular Medicine 2018, 96(10):983-992 (Review article). # - corresponding author
Mufazalov IA#, Kuschmann J, Andruszewski D, Masri J, Gabriel LA, Adams P, Reissig S, Hövelmeyer N and Waisman A#. Balanced Bcl-3 expression in murine CD4+ T cells is required for generation of encephalitogenic Th17 cells. European Journal of Immunology 2017, 47(8):1335-1341. # - corresponding author
Mufazalov IA#, Schelmbauer C, Regen T, Kuschmann J, Wanke F, Gabriel LA, Hauptmann J, Muller W, Pinteaux E, Kurschus FC and Waisman A#. IL-1 signaling is critical for expansion but not generation of autoreactive GM-CSF+ Th17 cells. The EMBO journal 2017, 36: p102-115. # - corresponding author
Mufazalov IA#, Regen T, Schelmbauer C, Kuschmann J, Muratova AM, Nikolaev A, Muller W, Pinteaux E and Waisman A#. Generation of a Novel T Cell Specific Interleukin-1 Receptor Type 1 Conditional Knock Out Mouse Reveals Intrinsic Defects in Survival, Expansion and Cytokine Production of CD4 T Cells. PLOS One 2016, 11: e0161505. # - corresponding author
Mufazalov IA# and Waisman A. Isolation of Central Nervous System (CNS) infiltrating cells. Methods in Molecular Biology 2016, 1304: p73-79 (Book chapter). # - corresponding author
AG Mufazalov
Alumni
PhD thesis
Carsten Schelmbauer (2018-2023)
Master thesis
Helena Schäfer (2023)
David Andruszewski (2019)
Laureen Gabriel (2015)
Bachelor thesis
Mirja Guskowski (2024, ongoing)
Leon Amelong (2024)
Katherine Junger (2023)
David Andruszewski (2018)
Carsten Schelmbauer (2014)
Janina Kuschmann (2012)
Part time technical assistant (HiWi)
Markus Schraft (2023)
Lena Scherer (2021-2023)
Patricia Winkler (2019-2021)
Internships
Kama Atretkhany, PhD student (2014, 2015, 2018)
Alisa Muratova, PhD student (2016)
