A new potent approach to treat steroid-refractory acute Graft-versus-Host Disease?

Clinical studies suggested that endothelial dysfunction and damage can be involved in the development and severity of acute graft-versus-host disease (aGvHD), a complication in patients undergoing allogeneic hematopoietic cell transplantation. In a new paper under the lead of Olaf Penack from the Charité Berlin we show extensive damage, structural changes, and dysfunction of the vasculature during aGvHD. Subsequently, therapeutic intervention with an already clinical approved endothelium-protecting agent improved outcome in a mouse model of steroid-refractory aGvHD.
Sildenafil steroid-refractory acute GvHD, GvH-GvL, Andreas Beilhack laboratory, Würzburg University Immunology Program, Immunotherapy research Germany

Sildenafil protects from endothelial damage in experimental steroid-refractory acute GvHD

Allogeneic hematopoietic stem transplantation (allo-HCT) is the only curative treatment option for many patients suffering from blood cancers. However, there are still risks for patients undergoing allo-HCT. One major complication is acute graft-versus-host disease (aGvHD) occuring in more than two thirds of patients. This inflammatory condition primarily affecting the skin, liver, and intestines. Although treatment with steroids is successful in most patients, about the 20%-25% of patients fail initial steroid treatment resulting in a very high mortality rate. Currently, no standard treatment for this steroid-refractory aGVHD is available and its pathobiology remains poorly understood, thereby hindering the development of novel therapeutic approaches.

The endothelium is the first contact for immunological effector cells in the blood and a key regulator in various inflammatory processes. The endothelium was shown to be relevant for early complications after allo-HCT such as transplantation-associated-microangiopathy, veno-occlusive disease, capillary leak syndrome and diffuse alveolar hemorrhage. Recent studies also suggested a critical role of the endothelium in aGvHD. Accordingly, we found increased percentage of apoptotic Casp3+ blood vessels in duodenal and colonic mucosa biopsies of patients with severe aGvHD. 

Light sheet fluorescence microscopy, LSFM, Andreas Beilhack laboratory Würzburg University Immunology Program

Pathologic vascular restructuring in intestinal acute GvHD

In mouse models of experimental aGvHD, we detected severe microstructural endothelial damage and reduced endothelial pericyte coverage accompanied by reduced expression of endothelial tight junction proteins leading to increased endothelial leakage in aGvHD target organs. Employing light-sheet fluorescence microscopy revealed structural changes in the colonic vasculature including increased vessel branching and vessel diameter. Human biopsies and murine tissues from steroid refractory aGvHD revealed extensive tissue damage but low levels of alloreactive T cell infiltration in target organs, providing the rationale for T-cell independent steroid refractory aGvHD treatment strategies. Consequently, we tested the endothelium-protective PDE5 inhibitor sildenafil, which reduced apoptosis and improved metabolic activity of endothelial cells in vitro. Accordingly, sildenafil treatment improved survival and reduced target organ damage during experimental steroid refractory aGvHD. The study by Steffen Cordes et al. demonstrates extensive damage, structural changes, and dysfunction of the vasculature during aGvHD. Consequently, therapeutic intervention by endothelium-protecting agents, such as sildenafil, appear attractive to treat steroid refractory aGvHD complementing current anti-inflammatory treatment options. 

 

This work resulted from a strong collaborative effort between clinicians and scientists from different European medical centers: The Charité Berlin and the University Hospitals of Barcelona (Spain), Hannover, Heidelberg and Würzburg. Our lab was supported for this study be the DFG collaborative research center TRR221 GvH-GvL (project B11).

 

Reference:

Cordes S, Mokhtari Z, Bartosova M, Mertlitz S, Riesner K, Shi Y, Mengwasser J, Kalupa M, McGeary A, Schleifenbaum J, Schrezenmeier J, Bullinger L, Diaz-Ricart M, Palomo M, Carrreras E, Beutel G, Schmitt CP, Beilhack A, Penack O. (2020). Endothelial damage and dysfunction in acute graft-versus-host disease. Haematologica, in press (doi: 10.3324/haematol.2020.253716).

Breaking a vicious cycle in multiple myeloma

Multiple myeloma is a malignant disease of antibody producing plasma cells. As this devastating type of cancer locates to the bones it seems that multiple myeloma highly depends on close interactions with the bone marrow microenvironment. In our new paper by clinician-scientist Dr. Antonio Solimando et al. we describe our new discovery that multiple myeloma cells use an adhesion molecule called JAM-A (Junctional-adhesion-molecule A) to interact with blood vessel lining endothelial cells. It appears as these endothelial-multiple myeloma interactions feed into a vicious cycle propagating disease progression.

Multiple myeloma, bone marrow environment, adhesion molecules, JAM-A, Andreas Beilhack laboratory, cancer immunotherapy, Antonio Solimando, vicious cycle, new treatment
Our projected started with our initial observation that the expression level of JAM-A by malignant plasma cells can predict disease outcome. Subsequently, we discovered that elevated membrane expression of JAM-A also on bone marrow endothelial cells of patients with newly diagnosed or relapsed-refractory multiple myeloma cells predicted poor clinical outcome.

Based on this finding we investigated how elevated JAM-A levels would contribute to more aggressive disease. We discovered that direct contact of endothelial cells with multiple myeloma cells would enhance JAM-A levels. Then it got even more interesting, as the cell adhesion molecule JAM-A has remarkable features: it can interact with itself if expressed on two opposing cell types. Furthermore, if JAM-A is shed by a cell, the soluble form of the JAM-A molecule can bind to cell-bound JAM-A, which in turn even enhances its binding capacity. What ensues is a vicious cycle of malignant plasma cells expressing and shedding JAM-A, increasing JAM-A expression on endothelial cells and stimulating blood vessel formation. In turn, increasing numbers of JAM-A-overexpressing endothelial cells can now better bind malignant plasma cells, which now find more interaction partners and by increasing the multiple myeloma niche space can produce more JAM-A. Consequently, using different experimental models we found that blocking the adhesion molecule JAM-A would inhibit blood vessel formation, reduce JAM-A interactions and impair multiple myeloma disease progression. These therapeutic effects of blocking JAM-A were observed in preclinical models not in patients and, therefore, must be interpreted with caution. Nevertheless, our new findings may point towards a potential Achilles’ heel of multiple myeloma that might be exploited therapeutically in the future.

This work was generally supported by the Bavarian Research Foundation within the research consortium FORTiTher and the German Research Council (DFG) consortium µbone.

Reference:

Solimando AG, Da Vià MC, Leone P, Borrelli P, Croci GA, Tabares P, Brandl A, Di Lernia G, Bianchi FP, Tafuri S, Steinbrunn T, Balduini A, Melaccio A, De Summa S, Argentiero A, Rauert-Wunderlich H, Frassanito MA, Ditonno P, Henke E, Klapper W, Ria R, Terragna C, Rasche L, Rosenwald A, Kortüm KM, Cavo M, Ribatti D, Racanelli V, Einsele H, Vacca A, Beilhack A. (2020). Halting the vicious cycle within the multiple myeloma ecosystem: blocking JAM-A on bone marrow endothelial cells restores the angiogenic homeostasis and suppresses tumor progression. Haematologica, in press

Nature Reviews Microbiology features our recent research paper

Nature Reviews Microbiology, Andreas Beilhack lab, host-pathogen interactions, Würzburg University, Germany, Imaging, Light-sheet fluorescence microscopy, immunotherapyIn its current issue, the journal Nature Reviews Microbiology reports about our recent mBio research article advancing light-sheet fluorescence microscopy (LSFM) to study host-pathogen interactions within the 3D environment of the lung. In the news section Under the Lens, Manish S. Kushwah and Stephen Thorpe from Oxford University highlight our research article by former postdoctoral fellow Dr. Jorge Amich, postdoctoral fellow Zeinab Mokhtari et al. in their report A clearer picture of microbial biogeography.

Amich J*, Mokhtari Z*, Strobel M, Vialetto E, Sheta D, Yu Y, Hartweg J, Kalleda N, Jarick KJ, Brede C, Jordán-Garrote AL, Thusek S, Schmiedgen K, Arslan B, Pinnecker J, Thornton CR, Gunzer M, Krappmann S, Einsele H, Heinze KG, Beilhack A. (2020). 3D light sheet fluorescence microscopy of lungs to dissect local host immune – Aspergillus fumigatusinteractions. mBio 11(1): e02752-19

T cells induce interferon-dependent cell cycle regulator pathways in cancer cells as a key immune mechanism to control cancer

A new paper in Nature Communications by Brenner et al. demonstrates in mice and in human patients that cancer control strictly requires the activation of tumour-intrinsic, senescence-inducing cell cycle regulators by the immune system to stably arrest those cancer cells that escape from eradication.
Cancer immunosurveillance, cancer immune senescence-signalling, immunotherapy, Andreas Beilhack research laboratory, immunology program Würzburg university

Mechanisms of cancer immunosurveillance. (A) Conventional and cancer immunotherapies can enhance direct cytolytic effector mechanisms of cytotoxic T cells and NK cells. (B) TNF and interferon released by Th1 T cells can induce and maintain cancer cell senescence. Mutations in the interferon-senescence-signalling pathway in tumor metastases can abolish this important mechanism and result in cancer progression.

Recent advances in cancer immunotherapy allow to efficiently unleash immune effector cells. Consequently, T cells, NK cells and macrophages can kill malignant cells throughout the body in patients with hematologic and solid cancers. Even if these therapies primarily aim to completely eradicate all cancer cells, often enough cancer cell killing remains incomplete and does not sufficiently and permanently control cancer. Moreover, the majority of cancer-related deaths do not result from the primary tumor. Instead, months or even years after initial therapy, cancer metastases arise from reawakened, dormant cancer cells that had been resistant to chemo-, radiation- or immunotherapies.

In a collaborative multidisciplinary effort lead by the team of Prof. Dr. Martin Röcken from Tübingen University and scientists from Tübingen and Würzburg University, we uncovered that IFN-γ/STAT1-dependent activation of the senescence-inducing cell cycle regulators p16Ink4a/p19Arf and p21Cip1 is required to keep cancer cells in a senescent state that had escaped immune cell mediated killing. Conversely, metastases that acquire mutations in the IFN-γ-senescence-inducing signalling pathways become resistant to immunotherapies and progress. In line with this, our study discovered that more than half of the metastases of patients not responding to immune checkpoint inhibitor therapy had at least one defect or genetic alterations in the IFN-dependent senescence-signalling pathway. Consequently, drugs that can re-invigorate the senescence-signalling pathways in cancer cells such as CDK4/6 inhibitors appear promising to be combined with cancer immunotherapy.

This work pinpointing key mechanisms required for protection against cancer cells that escape from cytotoxicity was supported by the German Research Council (Deutsche Forschungsgemeinschaft) in the research consortium FOR2314 Targeting therapeutic windows in essential cellular processes for tumor therapy and the Wilhelm Sander-Stiftung.

Optical lung scans of host-pathogen interactions

In a new publication that appeared today in the journal mBio we present light-sheet fluorescence microscopy that pinpoints critical immune cell populations responding to an invasive fungal lung infection.

Read more about our new research article on how a powerful microscopy technique helps scientists to find a needle in a haystack – only 2 micrometer thick fungal spores (50 times thinner than a hair) in an entire lung to learn about the ensuing immune response.

A gala event for cancer research

In an outstanding gala event, our joint research endeavor with the team of Prof. Harald Wajant was awarded to advance cancer immunotherapy.

Cancer research, immunotherapy, Andreas Beilhack laboratory, Würzburg Germany, Stiftung Forschung Hilft! Gabriele NelkenstockOn November 16, 2019, the foundation Forschung Hilft! invited to the new Hotel Melchior Park in Würzburg for a fabulous fund-raising gala evening “Art & Music & Magic.” At the same event, our research effort towards developing new concepts for cancer therapy was honored: The foundation “Forschung Hilft” selected our research project for its annual Großen Förderpreis worth 20.000 Euros. Receiving this award was a wonderful and highly motivating experience knowing that Ms. Gabriele Nelkenstock and her dedicated team of the foundation raised the funds for this cancer research award from patients and committed private donors.

The award will directly support one of our cutting-edge projects to reactivate the body’s immune response against cancer. We want to achieve this by newly designed antibody fusion proteins. The effect of these antibody fusion proteins only unfolds in the immediate vicinity of cancer cells, so that the body’s immune response is only activated there. The advantage of our therapy concept is that no immune cells need to be manipulated from the outside. Furthermore, disadvantages of previously clinically tested immune-activating antibodies can be minimized, such as side effect profiles or systemic effects.

The research groups of Prof. Andreas Beilhack and Prof. Harald Wajant have collaborated on the immune regulation of inflammatory diseases and cancer for more than 10 years. In this collaborations, they particularly focus on the molecular and cellular mechanisms of action of the tumor necrosis factor receptor superfamily. The molecular biological and biochemical expertise of Prof. Wajant’s group and the experience of Prof. Beilhack’s group in immunotherapy and cancer complement each other in an excellent way. The interdisciplinary team consists of experienced biologists, molecular biologists, biochemists, bioinformaticians, technical assistants and clinicians.

The Große Förderpreis of the Foundation “Forschung Hilft!” sets the first foundation stone to develop a new type of cancer therapy. Thank you!

Fifth Joint Meeting of IMIHM and the FungiNet consortium

Invasive Aspergillosis, Immunotherapy, diagnosis, A. fumigatus, host-pathogen-interactions, Andreas Beilhack research laboratory, Würzburg University, GermanyFrom June 26th-27th, 2018, the DFG Transregio 124 FungiNet Consortium convened in Würzburg to present and discuss the very latest research advances on the interactions of pathogenic fungi and the host immune system.

This meeting provided the opportunity especially for young pre- and post-docs of the FungiNet Consortium to share their very latest research data in own sessions prior to the 12th International Meeting for Invasive Mycoses in Haematologic Malignancies (IMIHM XII) organized by Prof. Jürgen Löffler. The topics covered aspects of immunology, microbiology, clinical mycology as well as basic science. Postdoctoral fellow Zeinab Mokhtari from the Beilhack lab presented the progress of two key research projects in the FungiNet consortium. The vivid discussions continued with internationally renowned experts in the field of fungal infections at the IMIHM XII, when they presented the most recent developments in fungal research covering topics from fungal recognition receptors and inflammasome signaling to clinical challenges based on the emergence of fungal multi-drug resistance.

Zeinab Mokhtari, Axel Brakhage, Transregio TRR 124, FungiNet, Andreas Beilhack IZKF Forschergruppe Würzburg

The FungiNet Research Consortium under the leadership of Prof. Axel Brakhage is a joint interdisciplinary endeavor of researchers from the Universities of Jena and Würzburg and funded by the Deutsche Forschungsgemeinschaft (DFG). In 2017 the DFG extended its support for this research network for the next 4 years.

Program and pictures of the 5th Joint Meeting of the DFG TRR 124 FungiNet consortium and IMIHM XII.

New publication in Frontiers in Immunology

Photoconversion of fluorescent proteins enriches the toolbox to study cell migration in vivo. In a new paper Katja Jarick (née Ottmüller) describes how this procedure can be used to track alloreactive T cells migrating to graft-versus-host disease target tissues. 

Katja Jarick, GvHD research, Beilhack lab, Würzburg University, GermanyDendra2 is a fluorescent protein with fantastic features. In its natural state it provides green fluorescence. However, if cells expressing this fluorescent protein are exposed to light of a particular wavelength, their green color switches to red. In our paper by Jarick et al. we describe how this trick can be used to study the fate of effector T cells in mice. Dendra2 expressing cells can be photoconverted within tissues in real-time. Once photoconverted, they can be spatiotemporally tracked based on their unique color signature. Strikingly, even if T cells divided up to 4 times, we were still able to identify this photoconverted cells. Photoswitching of other fluorescent proteins has been reported before. However, Dendra2 proved optimal for T cell tracking due to high fluorescence quantum yields and low phototoxicity. Employing this technique now opens new avenues to study the fate of tumor infiltrating immune cell populations, cancer metastasis, migration patterns of allreactive T cells or the dynamics and plasticity of immune cell subsets in different scenarios such as infection, inflammation and immunotolerance. Our work has been generously supported by the IZKF Würzburg, the EFRE Program of the European Union and the DFG Transregio Research Network TRR221

Reference:

Jarick KJ, Mokhtari Z, Scheller L, Hartweg J, Thusek S, Le DD, Ranecky M, Shaikh H, Qureischi M, Heinze KG, Beilhack A. (2018). Photoconversion of Alloreactive T Cells in Murine Peyer’s Patches During Acute Graft-Versus-Host Disease: Tracking the Homing Route of Highly Proliferative Cells In Vivo. Front. Immunol. 9:1468.

13th Annual Immunology Network Meeting Erlangen – Tübingen – Würzburg

Annual meeting immunology program Erlangen, Tübingen, Würzburg UniversityThe annual meeting of the immunology programs from Würzburg, Erlangen and Tübingen Universities took place in Obertrubach, Franconia. This year’s meeting was organized by Hans-Martin Jäck and his team from the University of Erlangen.

From June 18th-20th, 2018, the 13thnetwork meeting of research training groups in immunology from the Universities of Erlangen-Nuremberg, Tübingen, Würzburg was held in the Obertrubach, a pristine place in the Veldensteiner Forst with its characteristic 120 million years old rock and karst formations. For three days, graduate students and faculty members met to present and discuss their research projects in a relaxed atmosphere. Already in the past years, this meeting has proven as a great opportunity to share new developments and concepts and to foster scientific collaboration. Highlights of this year’s retreat were the keynote lectures by Dirk Brenner (Luxembourg) and Marco Herold (Melbourne). Marco Herold, who is holding now a faculty position in Australia at the Walter and Eliza Hall Institute, had graduated from Würzburg University and is a former member of the Würzburg immunology graduate program. Musga Qureischi and Tim Steinfatt from the Beilhack lab gave lectures about their progress and Julia Hartweg, Haroon Shaikh and Leni Strobel presented their thesis projects in the poster session. All PhD students of the Beilhack lab are members of the graduate program Immunomodulation, which is integrated in the Würzburg Graduate School of Life Sciences (GSLS). Graduate students of this program are required to give at least one poster and one scientific presentation at this annual immunology network meeting.

Immunology Community Germany, Andreas Beilhack Lab, Würzburg UniversityImmunology Meeting, Obertrubach, Germany

 

 

Scholarship for Dalia Sheta

The foundation “Help in the fight against cancer e.V.” awarded Dalia Sheta, a talented master student in the Beilhack lab with a scholarship of 3,000€ for the year 2018.

THilfe im Kampf gegen Krebs - Immuntherapie, Andreas Beilhack Forschungslabor, Würzburg, Deutschlandhis scholarship honors the great commitment of Ms. Sheta for research at the University Hospital Würzburg. In a ceremony on May 18, 2018, Prof. Dr. Georg Ertl, director of the Würzburg University Clinics, and Ms. Gabriele Nelkenstock, initiator and chair of the two foundations for cancer researchhanded over the award to Ms. Dalia Sheta. 

Scholarship, master thesis, immunotherapy, Andreas Beilhack lab, Würzburg, Germany

After the scholarship ceremony on 18 May this year: Gabriele Nelkenstock and Professor Georg Ertl as well as scholars Dina Kouhestani, Maria Geis and Dalia Sheta (front, from left to right). Back: Roland Sauer (Managing Director of the DJK Rimparer Wölfe), Professor Gernot Stuhler and Professor Andreas Beilhack. (Photo: University Hospital Würzburg)

 

“Patients suffering from immunodeficiency are at high risk of infections. Immunodeficiency can be triggered by the underlying disease such as cancer, by cancer therapy or by stem cell transplantation. In my research project, I investigate which cytokines, important molecular messengers, regulate immune cells in the lungs that protect against life-threatening infections caused by fungi, bacteria or viruses. If it is possible to specifically and preemptively strengthen the immune system at critical entry sites for pathogens such as our lungs, life-threatening complications in immunocompromised cancer patients can be prevented.”

Dalia Sheta, master student, Beilhack lab

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