Internship/Master project: Advanced Microscopy of Basal Inhibitory Signaling

Work Activities
Visualizing Basal Inhibitory Signaling in T Cells Using Advanced Microscopy
Are you curious about how immune cells make life-or-death decisions? Do you want to explore the molecular tug-of-war between activation and inhibition in real time? Join us in the Physics of Cellular Interactions (PCI) group, led by Dr. Kristina Ganzinger, to investigate how inhibitory immune receptors like PD-1 suppress T cell activation.

In this project, you will use cutting-edge microscopy techniques to visualize the molecular dynamics of immune signaling. You will explore how PD-1, a key inhibitory receptor, can suppress T cell signaling through basal activity, and how novel therapeutic molecules - called RIPR molecules - can reverse this basal inhibition and have more potent effects.
Understanding and proving the existence of basal PD-1 signaling is crucial, as it may explain why many cancer patients do not respond to current PD-1/PD-L1 checkpoint blockade therapies. By uncovering this hidden layer of immune suppression, your work could help guide the development of more effective treatments that potentiate T cell activity against tumors.

More About the Project
T cells are activated when their T cell receptors (TCRs) bind to peptide-MHC (pMHC) complexes, triggering a phosphorylation cascade involving kinases like LCK and adaptor proteins like Zap70. However, this activation can be suppressed by inhibitory receptors such as PD-1, which recruit phosphatases like SHP1/2 to dephosphorylate key signaling proteins.

Recent studies suggest that PD-1 may exert basal inhibitory activity, even in the absence of its ligand PD-L1. This project aims to test that hypothesis using supported lipid bilayers (SLBs) and single-molecule fluorescence microscopy. You will:

• Reconstitute TCR and PD-1 signaling of cells on SLBs functionalized with pMHC and PD-L1.
• Measure Zap70 and SHP2 activity using total internal fluorescent (TIRF) microscopy.
• Compare how RIPR molecules - which recruit the phosphatase CD45 to PD-1 - can suppress PD-1 activity and restore TCR signaling.

This work is part of the REPRESSIT consortium (https://www.repressit.eu/), a European collaboration developing next-generation immunotherapies that go beyond traditional checkpoint inhibitors.

What You Will Learn

1. Preparation of functionalized supported lipid bilayers (SLBs).
2. Use of TIRF and single-molecule microscopy to study protein dynamics.
3. Quantitative analysis of spatiotemporal signaling patterns.
4. Insight into the biophysics of immune regulation and therapeutic design.

Background Reading

1. Fernandes, R.A., et al. Immune receptor inhibition through enforced phosphatase recruitment. Nature (2020).
2. Davis, S., et al. The kinetic-segregation model: TCR triggering and beyond. Nat Immunol. (2006).
3. Wei, S.C., et al. Fundamental mechanisms of immune checkpoint blockade therapy. Cancer Discovery (2018).

Qualifications
You are currently enrolled in a Master’s program in (bio)physics, molecular biology, biomedical sciences, or a related field. You have a nationality of an EU-member state and/or you are a student at a Netherlands University. The internship has to be a mandatory part of your curriculum. We expect you to be available for at least 6 months, although longer is preferable.

Work environment
AMOLF is a part of NWO-I and initiate and performs leading fundamental research on the physics of complex forms of matter, and to create new functional materials, in partnership with academia and industry. The institute is located at Amsterdam Science Park and currently employs about 140 researchers and 80 support employees. www.amolf.nl

Physics of Cellular Interactions (Ganzinger) group – We are a supportive and social research group of about 8 PhD students and postdocs, which work together in small teams on various projects from immunological signalling to signalling in synthetic cells. The collaborative and social atmosphere includes to the other research groups at the AMOLF institute at the Science Park in Amsterdam. Our group has been making recent technical advances, including a new single-molecule tracking method based on DNA-PAINT that we recently developed (Nat Commun 2021, Nat Commun 2023), new in vitro assays (ACS Syn Biol 2021, ACS Omega 2024), and imaging approaches developed by our lab and others (JACS 2013, Nat Immun 2016, PNAS 2019), all to closely investigate the intricate molecular details of cellular signaling.

https://amolf.nl/research-groups/physics-of-cellular-interactions

Working conditions
At the start of the traineeship your trainee plan will be set out, in consultation with your AMOLF supervisor.

More information?
For further information about the position, please contact Gerard Castro-Linares g.castrolinares@amolf.nl or Kristina Ganzinger: k.ganzinger@amolf.nl.

Application
You can respond to this vacancy online via the button below.

Online screening may be part of the selection.

Diversity code
AMOLF is highly committed to an inclusive and diverse work environment: we want to develop talent and creativity by bringing together people from different backgrounds and cultures. We recruit and select on the basis of competencies and talents. We strongly encourage anyone with the right qualifications to apply for the vacancy, regardless of age, gender, origin, sexual orientation or physical ability.

AMOLF has won the NNV Diversity Award 2022, which is awarded every two years by the Netherlands Physical Society for demonstrating the most successful implementation of equality, diversity and inclusion (EDI).

Commercial activities in response to this ad are not appreciated.