From understanding to rational design of next-generation cancer therapies

Projektinformationen

subLETHAL

ID Finanzhilfevereinbarung: 101054921

DOI

10.3030/101054921

EK-Unterschriftsdatum 21 September 2022

Startdatum 1 Dezember 2022

Enddatum 30 November 2027

Finanziert unter

• European Research Council (ERC)

Gesamtkosten

€ 2 499 893,00

EU-Beitrag

€ 2 499 893,00

Koordiniert durch

STICHTING RADBOUD UNIVERSITAIR MEDISCH CENTRUM
Netherlands

Periodic Reporting for period 1 - subLETHAL (From understanding to rational design of next-generation cancer therapies)

Berichtszeitraum: 2022-12-01 bis 2025-05-31

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

Immunotherapy combined with cytotoxic and molecular therapies have entered centre stage as novel treatments for cancer. Despite this advance, toxicity and insufficient efficacy in many patients remain major obstacles and illustrate the need for therapy improvement based on a deeper understanding of the cellular damage and repair responses to drugs and immunotherapy.
To overcome this roadblock, we have performed high-content screening of drug effects at high and low dose in combination with cytotoxic effecter cells and identified compounds and conditions which sensitize tumor cells to CTL-mediated cytotoxicity without compromising CTL viability, activation and proliferation. An innovative microscopy pipeline was developed to detect therapy stress at single cell level over time and quantify drug effects at high but also very low dose. Single-cell analyses are combined with advanced statistics and mathematical modeling to derive the damage profile and duration of damage for each modality in tumor cells. In conclusion, sublethal damage forms the basis of cytotoxic and immunotherapy and can be exploited for design of multi-targeted ultra-low dose additive regimens.

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

3D coculture models were used to screen cytostatic compounds and 50 small molecule inhibitors followed by principal component analysis to identify conditions which enhancing CTL-mediated killing of tumor cells without and toxicity of compounds on CTL. Key compounds were then investigated using high-content microscopy and automated image analysis for induction of sublethal damages, including membrane damage, DNA damage and intracellular reactive oxygen species (ROS). A subset of compounds was pursued for developing in silico models of drug effects and to build individual “damage fingerprints” to decompose the modes of action for each effective compound at sublethal dose. This allowed the hybrid continuous-discrete simulation of intracellular damage types and reactive cell responses, including cell aggregation and damage repair. To map molecular responses to sublethal damage induction, RNA sequencing was used to revealed a graded responses to sublethal and lethal doses of drugs. The data indicate that even at sublethal dose, with cell viability and morphology preserved, significant transcriptional changes and regulated pathways are engaged to secure cell homeostasis.

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

The technical advances include first-in-field imaging approach to record sublethal and lethal damages during immune-cell based and other anticancer therapies and tracking of multiple damage types, their time-resolved behaviors and cell-fate (survival, death) as an integrated process by advanced time-series data analysis methodology and mathematical modeling. The automated image analysis using AI and on-the-fly reporting delivers first-in-field to follow different cell damages during immunotherapy and treatment with drugs. The approach is generic and will be useful in a range of other settings, including monitoring phototoxicity during live-cell microscopy and cell stress during cancer invasion. The analysis of combined effects of compounds on tumor cell death and CTL activation and growth defines a much-needed workflow to deliver an integrated view on compound efficacy and safety. The in-silico models will be inferred from cell damage time course data. Here, wet-lab research combined with modeling will allow to predict drug effects at lethal and sublethal dose, which will advance predictions for meaningful combinations. In addition, novel 3- and 4-photon microscopy was introduced to the field of tumor immunotherapy in a preclinical mouse model.