17th European Molecular Imaging Meeting (EMIM) 2022

17th European Molecular Imaging Meeting (EMIM) 2022

March 15–18, 2022 | Thessaloniki, Greece

Understand nature’s complexity with cutting-edge imaging and microscopy technologies

We are excited to see you in Thessaloniki!

Join us for EMIM, the annual meeting of the European Society for Molecular Imaging (ESMI), to meet and exchange with experts from all fields of imaging science. Enlarge your network, learn more about state-of-the-art research and technological innovations, and be part of this vivid community.  

We would love to see you at our booth to explore our novel imaging technologies that will allow you to dive deep into your sample material. Check out the resources on this web page as a sneak peek of what you can expect to discuss with us in Greece. 

Helexpo Congress Centre
Thessaloniki, Greece
Miltenyi Biotec Booth #107
March 15–18, 2022
 

Scientific contribution

Scientific talk

Helexpo Congress Centre
Thessaloniki, Greece
Room 01
Parallel session, PS 21-03
March 18, 2022 09:30–9:40 a.m.
 
In vivo tracking of bioluminescent CAR T cells in a pancreatic adenocarcinoma xenograft model reveals anti-tumor response via antigen-recognition dependent activation and expansion of therapeutic cells  

Presenter:  Katharina Wittich

Katharina Wittich, Olaf Hardt, Rita Pfeifer 
Miltenyi Biotec B.V. & CO. KG, R&D Reagents, Bergisch Gladbach, North Rhine-Westphalia, Germany 
 

Abstract

INTRODUCTION
Chimeric antigen receptor (CAR) T cells and their therapeutic success in hematological cancer patients has received growing attention in oncology research. Despite such striking clinical success in hematological malignancies, CAR T therapies have failed to reproduce this efficacy in solid tumors. Development of future immunotherapies requires deeper understanding in the behavior of CAR T cells. In vivo CAR T cell tracking can provide useful spatiotemporal information and allows longitudinal monitoring of T cell trafficking, in vivo proliferation, persistence and activation state.

METHODS
CAR constructs were modified for transgenic T cells to show constitutive CAR expression and simultaneously encode for a click beetle luciferase (CBR2) under the control of an inducible NFAT-AP/1 promoter. CARs were directed either against an antigen which was expressed by the tumor cells (inducible EGFR-CAR, therapy group) or a non-expressed antigen (inducible BDCA2-CAR, mock control). The inducible system enables T cell activation dependent luciferase expression. Activated CAR positive T cells were monitored longitudinally using in vivo bioluminescence imaging (BLI) in a subcutaneous pancreatic adenocarcinoma xenograft model. Concurrent detection of tumor burden was achieved by 2D fluorescence imaging (FLI) of an endogenously expressed reporter protein (TurboRFP). 

RESULTS
Longitudinal tracking of activated CAR T cells in vivo revealed antigen recognition dependent intra-tumoral CAR T cell activation and expansion. In the therapy group, we could detect early accumulation of activated transgenic T cells at the tumor site compared to mock control. An increase in bioluminescence signal over time indicated antigen dependent proliferation and expansion of activated CAR+ T cells. Additionally, we could show a decrease in fluorescent signal of TurboRFP expressing tumor cells, which was not replicated in the cohort receiving mock control CAR T cells. Both cohorts, untreated and mock control, showed continuous tumor growth with tumor sizes reaching the abort criteria. However, reduction of tumor burden shows temporal correlation with a peak in bioluminescence signal of activated transgenic T cells in the tumor of the therapy group. All in all, we established a non-invasive, inducible tracking system, in order to trace activated CAR T cells in vivo.

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Scientific poster

Helexpo Congress Centre
Thessaloniki, Greece
Exhibition Hall 
Poster session 1
March 16, 2022 7:15 p.m.–9:00 p.m.
 
Multi-modal imaging workflow for monitoring CAR T cell therapy against solid tumors from whole-body to single-cell level 

Presenter:  Rita Pfeifer

Rita Pfeifer, Katharina Wittich, Cathrin Linnartz, Olaf Hardt 
Miltenyi Biotec B.V. & CO. KG, R&D Reagents, Bergisch Gladbach, North Rhine-Westphalia, Germany
 

Abstract

INTRODUCTION
As CAR T cell therapy continues to become a mainstay pillar in cancer treatment, there is a growing need to understand the function and interaction of such therapeutic cells in their native environment to further improve treatment modality. Imaging platforms with spatiotemporal information can provide profound insight into therapeutic ongoings during different therapy stages, accelerating the establishment of successful therapies. Thus, we aimed to retrieve multi-scale information on therapeutic effects following CAR T cell therapy using various imaging approaches. 

METHODS
In vivo and ex vivo CAR T cell tracking was conducted using a subcutaneous pancreatic adenocarcinoma xenograft model. For this, T cells were genetically equipped with a bicistronic transgene to express the CAR of interest as well the CBR2 luciferase. With the start of therapy, longitudinal whole-body imaging was performed by means of bioluminescent and fluorescent tomography (BLT/FLT). At selected time points, tumors were removed and subjected to high-resolution ex vivo analysis of CAR T cell infiltration using 3D light sheet fluorescence microscopy (LSFM, Ultramicroscope Blaze™). Subsequent, deep-phenotyping of intratumoral T cells was conducted by cyclic immunofluorescence staining (MICS, MACSima™ Imaging Platform).

RESULTS
Longitudinal whole-body in vivo BLT/FLT imaging displayed differential bio-distribution of therapeutic and mock CAR T cells, in addition to a tumor antigen-dependent accumulation of immune cells at the lesion site. Notably, T cells homing and accumulation within the tumor did not correlate with therapeutic efficacy throughout the study suggesting a complex interplay between the CAR T and tumor cells. Subsequent high-resolution profiling of tumors by LSFM revealed the majority of tumor-infiltrating T cells were located within the tumor periphery in a heterogeneous, island-like distribution with only a small fraction reaching the tumor core. Moreover, the peripheral T cell clusters displayed a higher density than those in the inner tumor layers, suggesting a more pronounced anti-tumor response in the periphery. An in-depth characterization of the intratumoral T cells by means of MICS imaging revealed phenotypic differences between CAR and mock T cells. 

CONCLUSION
In this study we describe a multi-modal imaging workflow that allows the analysis of CAR T cell therapies at different scales ranging from whole-body to single-cell level. This workflow facilitates the investigation of CAR T cells in vivo and provides new insights for the development of novel, more efficacious CAR T cell immunotherapies.

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Technologies

Discover how ultra high content imaging, large-scale 3D light sheet microscopy, and multiphoton microscopy can open new avenues in cancer and immuno-oncology research.  

 

Applications and publications

Laying the foundation to understand tumor microenvironment (TME) complexity. See how researchers use technologies like ultra-high content imaging or 3D visualization by light sheet microscopy to dive deep into limited sample material and define new targets for immunotherapies.

 

Webinars and event recordings

Food for thought needed? Tune in to our webinars or watch the recordings of previous events such as the MACS® Imaging and Microscopy Day. Leading scientists from the fields of immuno-oncology and neuroscience presented recent findings achieved through groundbreaking imaging and microscopy technologies. 

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