Periodic Reporting for period 4 - H I C I (Transcriptional and epigenetic control of tissue regenerative HB-EGF in autoimmune CNS inflammation)
Período documentado: 2023-09-01 hasta 2025-02-28
Multiple Sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system (CNS), which affects young individuals during the most productive phases of their lives. In 85% of the patients, MS initially presents with a relapsing-remitting clinical course (relapsing-remitting MS, RRMS), which in the majority of these individuals is followed by a progressive stage with irreversible accumulation of neurologic deficits (secondary progressive MS, SPMS). A subset of patients develops a primary progressive variant of MS (PPMS), in which chronic accumulation of disability without super-imposed relapses is the most prominent clinical feature. Especially for chronic stages, only limited therapeutic options are available to date.
Moreover, in recent years both incidence and prevalence of MS have increased for yet unknown reasons, strengthening the need for efficacious therapies targeting acute and progressive stages in MS. Indeed, in spite of treatment strategies available already, the majority of MS patients will develop relevant neurological deficits over the course of the disease. As such, recent studies have shown that about 30-50% of MS patients will develop walking impairment over the course of 25 years of MS. Given that the age of onset in MS is between 20 and 30 years of age in most patients, this poses a relevant burden and threat to MS patients and their families, but also society in general. In particular, our understanding of the underlying pathology in progressive stages of MS is still limited, which has also hampered the development of novel therapeutics to causally influence these stages of the disease.
Given the increasing incidence of MS and our limited therapeutic options for progressive MS stages outlined above, developing novel strategies to tackle acute and particularly chronic MS stages in order to prevent disease progression is of utmost importance both for individual patients as well as for society, since available treatments, rehabilitation costs as well as the risk of unemployment pose relevant risks both for patients and society.
From a pathophysiological standpoint, local anti-inflammatory and regenerative mechanisms limit the acute inflammatory processes and define the extent of tissue recovery and consecutive residual neurological deficits both during acute and progressive disease stages, but fail over time causing progressive disease worsening. Astrocytes are CNS resident cells with important roles during both stages of MS. Their actions can either favor chronification or promote resolution of the inflammatory processes. We and others have identified several mechanisms, by which secreted factors from astrocytes promote the ongoing immune response. Even though astrocytes also secrete tissue-regenerative factors such as the leukemia inhibitory factor (LIF) or epidermal growth factor (EGF), the translation of these factors into translationally relevant strategies has been challenging due to an incomplete understanding of their cellular targets and molecular regulation. Thus, the discovery of novel tissue-protective factors and the understanding of their regulation in acute and chronic stages of MS yields the potential to develop novel therapies for chronic inflammatory and degenerative diseases of the CNS.
In this context, we have discovered a novel potential protective factor secreted from astrocytes, the so-called Heparin-binding EGF-like growth factor (HB-EGF). Our preliminary experiments have determined that HB-EGF is efficient in the initial stages to limit autoimmune CNS inflammation, but fails over time due to epigenetic alterations in its promoter region, which limit its availability and function. This process might contribute to disease progression, which makes HB-EGF and its regulation a potential treatment target in MS.
Our overall aim within this project therefore is to (i) examine role and (ii) regulation of HB-EGF in acute and progressive stages of autoimmune CNS inflammation for tissue recovery. In a more translational part of the project, we next aim to determine (iii) the potential therapeutic value of HB-EGF and its epigenetic regulation in acute and chronic autoimmune inflammatory CNS diseases. Finally, we aim to (iv) determine and its relevance as novel biomarker in MS, which might help to stratify patients at risk for disease progression.
Moreover, in recent years both incidence and prevalence of MS have increased for yet unknown reasons, strengthening the need for efficacious therapies targeting acute and progressive stages in MS. Indeed, in spite of treatment strategies available already, the majority of MS patients will develop relevant neurological deficits over the course of the disease. As such, recent studies have shown that about 30-50% of MS patients will develop walking impairment over the course of 25 years of MS. Given that the age of onset in MS is between 20 and 30 years of age in most patients, this poses a relevant burden and threat to MS patients and their families, but also society in general. In particular, our understanding of the underlying pathology in progressive stages of MS is still limited, which has also hampered the development of novel therapeutics to causally influence these stages of the disease.
Given the increasing incidence of MS and our limited therapeutic options for progressive MS stages outlined above, developing novel strategies to tackle acute and particularly chronic MS stages in order to prevent disease progression is of utmost importance both for individual patients as well as for society, since available treatments, rehabilitation costs as well as the risk of unemployment pose relevant risks both for patients and society.
From a pathophysiological standpoint, local anti-inflammatory and regenerative mechanisms limit the acute inflammatory processes and define the extent of tissue recovery and consecutive residual neurological deficits both during acute and progressive disease stages, but fail over time causing progressive disease worsening. Astrocytes are CNS resident cells with important roles during both stages of MS. Their actions can either favor chronification or promote resolution of the inflammatory processes. We and others have identified several mechanisms, by which secreted factors from astrocytes promote the ongoing immune response. Even though astrocytes also secrete tissue-regenerative factors such as the leukemia inhibitory factor (LIF) or epidermal growth factor (EGF), the translation of these factors into translationally relevant strategies has been challenging due to an incomplete understanding of their cellular targets and molecular regulation. Thus, the discovery of novel tissue-protective factors and the understanding of their regulation in acute and chronic stages of MS yields the potential to develop novel therapies for chronic inflammatory and degenerative diseases of the CNS.
In this context, we have discovered a novel potential protective factor secreted from astrocytes, the so-called Heparin-binding EGF-like growth factor (HB-EGF). Our preliminary experiments have determined that HB-EGF is efficient in the initial stages to limit autoimmune CNS inflammation, but fails over time due to epigenetic alterations in its promoter region, which limit its availability and function. This process might contribute to disease progression, which makes HB-EGF and its regulation a potential treatment target in MS.
Our overall aim within this project therefore is to (i) examine role and (ii) regulation of HB-EGF in acute and progressive stages of autoimmune CNS inflammation for tissue recovery. In a more translational part of the project, we next aim to determine (iii) the potential therapeutic value of HB-EGF and its epigenetic regulation in acute and chronic autoimmune inflammatory CNS diseases. Finally, we aim to (iv) determine and its relevance as novel biomarker in MS, which might help to stratify patients at risk for disease progression.
The project HICI : Transcriptional and epigenetic control of tissue regenerative HB-EGF in autoimmune CNS inflammation was started at the Department of Neurology at Technical University in Munich in March 2020, where a PhD student was hired. After accepting an appointment as Full Professor of Neuroimmunology (W3) at the Department of Neurology at Friedrich-Alexander Universtiy Erlangen Nuernberg, we transitioned to Erlangen together with the student starting on 1st of September 2020.
Here, we set up the laboratory, technical and legal requirements and started to perform and continue on the experiments focussing on Aims 1, 2 and 3 after transferring the grant to FAU Erlangen Nuernberg. Even though this process took several months and was complicated by the limitations and restrictions of the SARS-CoV2 pandemic, we were able to continue with the experiments planned for Aims 1, 2 and 3.
Indeed, we performed in vivo experiments outlined in Aim 1 knocking-down and analyzing the effects of astrocytic Hbegf during experimental autoimmune CNS-inflammation. In Aim 2, we performed fine-tuned experiments to dissect transcriptional and epigenetic regulation of HB-EGF in astrocytes. We were able to determine transcriptional and epigenetic regulation of HB-EGF and set up epigenetic analyses including bisulfite sequencing. Moreover, treatment experiments outlined in Aim 3 using HB-EGF and epigenetic modification were initialized as well and are currently ongoing together with further mechanistic studies on epigenetic modulation of HB-EGF. In Aim 4, we performed analyses on samples from MS patients and controls, which pointed towards a potential role of HB-EGF over the course of the disease. Our current analyses are focusing on the role of HB-EGF as well as th HB-EGF promoter methylation status as novel biomarkers for disease course, severity, and progression risk in acute and progressive stages of MS.
Most of these experiments led to meaningful results, pointing towards a functional and therapeutical relevance of HB-EGF and its regulation in autoimmune CNS inflammation. In that way, within the time period covered by this reporting period, we were able to successfully transition from Munich to Erlangen, transfer the grant, acquire laboratory and animal facility authorizations and perform major experiments described in the grant. We are now seeking to hire a postdoctoral fellow to be involved in the project. Together, major steps and milestones were achieved, which now are pursued further. Together, we are confident to further refine the understanding of HB-EGF biology and its relevance in MS within the upcoming period following this report.
Here, we set up the laboratory, technical and legal requirements and started to perform and continue on the experiments focussing on Aims 1, 2 and 3 after transferring the grant to FAU Erlangen Nuernberg. Even though this process took several months and was complicated by the limitations and restrictions of the SARS-CoV2 pandemic, we were able to continue with the experiments planned for Aims 1, 2 and 3.
Indeed, we performed in vivo experiments outlined in Aim 1 knocking-down and analyzing the effects of astrocytic Hbegf during experimental autoimmune CNS-inflammation. In Aim 2, we performed fine-tuned experiments to dissect transcriptional and epigenetic regulation of HB-EGF in astrocytes. We were able to determine transcriptional and epigenetic regulation of HB-EGF and set up epigenetic analyses including bisulfite sequencing. Moreover, treatment experiments outlined in Aim 3 using HB-EGF and epigenetic modification were initialized as well and are currently ongoing together with further mechanistic studies on epigenetic modulation of HB-EGF. In Aim 4, we performed analyses on samples from MS patients and controls, which pointed towards a potential role of HB-EGF over the course of the disease. Our current analyses are focusing on the role of HB-EGF as well as th HB-EGF promoter methylation status as novel biomarkers for disease course, severity, and progression risk in acute and progressive stages of MS.
Most of these experiments led to meaningful results, pointing towards a functional and therapeutical relevance of HB-EGF and its regulation in autoimmune CNS inflammation. In that way, within the time period covered by this reporting period, we were able to successfully transition from Munich to Erlangen, transfer the grant, acquire laboratory and animal facility authorizations and perform major experiments described in the grant. We are now seeking to hire a postdoctoral fellow to be involved in the project. Together, major steps and milestones were achieved, which now are pursued further. Together, we are confident to further refine the understanding of HB-EGF biology and its relevance in MS within the upcoming period following this report.
As outlined above, we were able to perform a relevant part of the experiments planned for Aims 1, 2, 3 and 4.
Indeed, we performed in vivo experiments outlined in Aim 1 knocking-down and analyzing the effects of astrocytic Hbegf during experimental autoimmune CNS-inflammation. We were able to determine a tissue-protective and regenerative function of HB-EGF during experimental autoimmune CNS-inflammation due to its effect on infiltrating immune cells, glial cells, as well as tissue-resident and infiltrating macrophages. Importantly, our analyses detected a myelination-stimulating effect of HB-EGF, which might be of particular relevance for acute and progressive stages of MS.
In Aim 2, we performed fine-tuned experiments to dissect transcriptional and epigenetic regulation of HB-EGF in astrocytes. We were able to determine transcriptional and epigenetic regulation of HB-EGF and set up epigenetic analyses including bisulfite sequencing. These studies were able to dissect a novel mechanism, by which certain transcription factors compete for their binding at the Hbegf promoter, a mechanism of potential interest not only to MS, but also hypoxic or degenerative CNS disorders.
Moreover, treatment experiments outlined in Aim 3 using HB-EGF and epigenetic modifiers were performed and pointed towards a therapeutic potential of HB-EGF and its epigenetic regulation in acute and chronic stages of experimental autoimmune CNS-inflammation. Next, mechanistic studies on epigenetic modulation of HB-EGF are currently ongoing and will complement our insight into these potentially druggable mechanisms.
In Aim 4, we performed analyses on samples from MS patients and controls, which pointed towards a potential role of HB-EGF over the course of the disease. Our current analyses are focusing on the role of HB-EGF as well as th HB-EGF promoter methylation status as novel biomarkers for disease course, severity, and progression risk in acute and progressive stages of MS.
Our ongoing experiments are fine-dissecting these main results on organismal, cellular, transcriptional and epigenetic level to complement our current results. Together, the available data and the datasets to be acquired within the next period might contribute to a deeper understanding of the role and regulation of HB-EGF in MS and point towards novel treatment targets in relapsing-remitting, secondary and primary progressive MS.
Indeed, we performed in vivo experiments outlined in Aim 1 knocking-down and analyzing the effects of astrocytic Hbegf during experimental autoimmune CNS-inflammation. We were able to determine a tissue-protective and regenerative function of HB-EGF during experimental autoimmune CNS-inflammation due to its effect on infiltrating immune cells, glial cells, as well as tissue-resident and infiltrating macrophages. Importantly, our analyses detected a myelination-stimulating effect of HB-EGF, which might be of particular relevance for acute and progressive stages of MS.
In Aim 2, we performed fine-tuned experiments to dissect transcriptional and epigenetic regulation of HB-EGF in astrocytes. We were able to determine transcriptional and epigenetic regulation of HB-EGF and set up epigenetic analyses including bisulfite sequencing. These studies were able to dissect a novel mechanism, by which certain transcription factors compete for their binding at the Hbegf promoter, a mechanism of potential interest not only to MS, but also hypoxic or degenerative CNS disorders.
Moreover, treatment experiments outlined in Aim 3 using HB-EGF and epigenetic modifiers were performed and pointed towards a therapeutic potential of HB-EGF and its epigenetic regulation in acute and chronic stages of experimental autoimmune CNS-inflammation. Next, mechanistic studies on epigenetic modulation of HB-EGF are currently ongoing and will complement our insight into these potentially druggable mechanisms.
In Aim 4, we performed analyses on samples from MS patients and controls, which pointed towards a potential role of HB-EGF over the course of the disease. Our current analyses are focusing on the role of HB-EGF as well as th HB-EGF promoter methylation status as novel biomarkers for disease course, severity, and progression risk in acute and progressive stages of MS.
Our ongoing experiments are fine-dissecting these main results on organismal, cellular, transcriptional and epigenetic level to complement our current results. Together, the available data and the datasets to be acquired within the next period might contribute to a deeper understanding of the role and regulation of HB-EGF in MS and point towards novel treatment targets in relapsing-remitting, secondary and primary progressive MS.