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Personalized Cancer Medicine is rapidly developing field that includes predictive medicine, preventive medicine and various personalized or individualized therapies, e.g. labeled “precision medicine”. One particular challenge with cancer is that origin of each cancer is a clonal event evolving into tumor heterogeneity. We focus on Chronic Lymphocytic Leukemia (CLL), Multiple Myeloma and Follicular lymphoma (FL) that are currently considered incurable. Although current treatment regimens prolong life for patients, CLL and MM cancer eventually relapse. Current challenges in using therapeutics against CLL and MM includes design of optimal treatment for individual patients based on characterization the tumor and its intratumor heterogeneity as observed by whole genome sequencing. Efficient therapies require a personalized approach that combines targeting lymphoma cells and the tumor microenvironment by restoring the patient’s own anti-tumor immunity.

One solution to this challenge is the so-called “n-of-one” studies where protocols are organized with diagnostically based patient stratification to individualized treatments (n=1).

To introduce individualized treatment for patients against available therapies, we aim to established cell-based assays and drug sensitivity platform at NCMM, University of Oslo and Oslo University Hospital. To establish a pipeline for direct drug senstivity screening in CLL and MM (WP1-Path A). To Complement the results from WP1-Path A, with Signaling pathway analysis (WP2-Path B) towards testing in xenografted mice and implementing therapy in n-of-one clinical trials. To Offer patients with intractable CLL and MM individualized treatment with an effective combination of targeted therapies.

We culture CLL cells with combination of feeder cells that express CD40L, APRIL and BAFF for 24 hours stimulation

We perform drug sensitivity screening with Prestimulated CLL cells in 384 well formats without feeder cells.

We culture MM cells in 384 well format for drug screening in response to T_helper cells prestimulation in the presence of IL2.

To support high-throughput drug sensitivity screening, We use cell-based assays such as CellTiter-Glo® Luminescent Cell Viability Assay and CellTox™ Green Cytotoxicity Assay to define drugs that inhibit cancer cell growth.

Additional methods such as cell proliferation assay, CellTox Green, apoptosis and oxidative stress (glutathione release) are also applied.

We also use established cell barcoding on CLL/MM for flow cytometry (7-AAD/BrdU cell proliferation and Caspase8/9 apoptosis assay).

Time course measurement using cell proliferation, CellTox-Green assay for CLL cells (unstimulated and soluble CD40 ligand stimulated) has been performed.

Time course measurement (0,24,48,72 hrs and 5 days) using cell proliferation, CellTox-Green assay for M2 cells has been performed. Benzalkonium chloride (BzCl) is used as Positive control.

Endpoint measurement using CellTiter-Glo assay for CLL and MM cells was performed with cell density of 5000.

Dose Response curve for 50 drugs has been generated for CLL patients (n=4) and MM (n=4).

We perform drug sensitivity screening to select potential drug candidates and pathway inhibitors through an approach where we directly assess patient samples. Selected drug candidates will first be validated by bioassays and flow cytometry to assess effects on intracellular mitogenic pathways (phosphoflow-based approach). We propose to use the drug sensitivity screening platform to identify and validate drug candidates for xenografting and “n-of-one” clinical trial studies.