Friday, 12 April 2024 11:00am – 12:00pm

This seminar will be delivered in Lecture Theatre 4

Speaker: Dr May C. Morris

Host: Dr Constance Bailey

Title: Designing Smart Tools and Strategies to Profile and Target Protein Kinases in Human Cancers: Fluorescent biosensors and their applications for diagnostics and drug discovery

Abstract: Protein kinases (PK) are hyperactivated in many human cancers thereby constituting relevant biomarkers and attractive pharmacological targets (Fleuren et al. 2016; Wu et al. 2015 ; Wu et al. 2016 ; Roskoski R.Jr 2021; Cohen et al. 2021). Although PK biomarkers may be detected through antigenic, proteomic, transcriptomic or genetic approaches, quantification of their expression does not convey an accurate readout of their activity, since they are subject to multiple posttranslational regulations. There are however currently no approaches that report on their functional activity for diagnostic purposes. In order to monitor PK activities in complex biological samples, we have developed a toolbox of fluorescent biosensors through conjugation of environmentally-sensitive fluorescent probes to peptide scaffolds derived from natural substrates (Morris M.C. 2022a & b). These synthetic biosensors enable quantification of PK activities in vitro, in cellulo and in vivo and can report on therapeutic inhibition in a sensitive and selective fashion (Prével C. et al. 2016; Gonzalez-Vera et al. 2017; Henri et al. 2019, Soamalala et al. 2020, Peyressatre et al. 2020, Tilmaciu et al. 2021). We have further combined our ensemble of fluorescent biosensors to profile several PK activities simultaneously in human biopsies and have established a multiplex optical biosensing approach that highlights distinctive signatures in lung adenocarcinoma, lymphoma and pancreatic cancer, thereby providing relevant functional information diagnostic purposes (Royet et al.in revision).

Moreover although a large number of inhibitors targeting the ATP-pocket of PKs have been approved by the FDA, they suffer limitations, including poor selectivity and emergence of resistance (Wu et al. 2015; Wu et al. 2016; Wilson et al. 2018). Efforts to develop new classes of drugs targeting the conformational plasticity or essential interactions between PKs and their partners are believed to offer promising perspectives for more selective therapeutics (Tong & Seeliger 2015). With the aim of targeting the conformational activation of CDKs, we have developed a fluorescent conformational biosensor technology that discriminates against ATP-pocket binding inhibitors, which we have successfully implemented to screen several libraries of small molecules thereby identifying original allosteric inhibitors (Prével et al. 2014a & b, Pellerano et al. 2017, Peyressatre et al. 2020). Lead compounds are new and druggable chemical scaffolds that efficiently inhibit cancer cell proliferation and migration through original and unexpected mechanisms of action which we have inestigated through combination of biochemical and imaging approaches. Our studies highlight the relevance of rational design strategies to engineer conformational biosensors for HTS programmes, as an alternative to conventional activity-based assays, enabling identification of new generations of small molecule allosteric kinase inhibitors for anticancer therapeutics.

References

Cohen P et al. Kinase drug discovery 20 years after imatinib: progres and future directions. Nat Rev Drug Discov. 2021; 20: 551-569.

Fleuren EDG et al.. The kinome ‘at large’ in cancer. Nat Rev Cancer. 2016; 16: 83–98.

Gonzalez Vera JA., et al. Lanthanide-based peptide biosensor to monitor CDK4/cyclin D kinase activity. Chem. Commun. 2017; 53: 6109-6112.

Henri P, et al. Psoriatic epidermis is associated with upregulation of CDK2 and inhibition of CDK4 activity. Br J Dermatol. 2020; 182: 678-689.

Morris MC. Shining Light on Protein Kinase Biomarkers with Fluorescent Peptide Biosensors. Life. 2022; 12: 516. https://doi.org/10.3390/life12040516

Morris MC. A Toolbox of Fluorescent Peptide Biosensors to Highlight Protein Kinases in Complex Samples: Focus on Cyclin-Dependant Kinases. European J Organic Chem. 2022; https://doi.org/10.1002/ejoc.202200120R1

Peyressatre M, et al. Fluorescent Biosensor of CDK5 Kinase ctivity in Glioblastoma Cell Extracts and Living Cells. Biotechnol. J. 2020; 15: 1900474.

Peyressatre M. et al. Identification of Quinazolinone Analogs Targeting CDK5 Kinase Activity and Glioblastoma Cell Proliferation. Frontiers Chem. 2020, 8, 691

Prével C, et al. Fluorescent peptide biosensors for monitoring CDK4/cyclin D kinase activity in melanoma cell extracts, mouse xenografts and skin biopsies. Biosensors and Bioelectronics. 2016; 85: 371-380.

Prével, C., et al. Fluorescent Biosensors for High Throughput Screening of Protein Kinase Inhibitors. Biotechnology J. 2014, 9, 253-65

Prével C. et al. Fluorescent biosensors for drug discovery- New tools for Old Targets – Screening for Inhibitors of Cyclin-dependent kinases. European Journal of Medicinal Chemistry. 2014, 88, 74-88

Roskoski R. Properties of FDA-approved smallvmolecule protein kinase inhibitors: A 2021 update. Pharmacological Research. 2021; 165: 105463.

Royet C, et al. Profiling CDK kinase activities in tumour biopsies through mutiplexed biosensing with fluorescent peptides. ACS Sensors, in revision

Soamalala J, et al. Fluorescent Peptide Biosensor for probing CDK6 Kinase Activity in Lung Cancer Cell Extracts. ChemBioChem. 2021; 22: 1065-1071.

Tîlmaciu CM., et al. Nanobiosensor Reports on CDK1 Kinase Activity in Tumor Xenografts in Mice. Small. 2021; 17: 2007177.

Tong & Seeliger. Targeting conformational plasticity of protein kinases. ACS Chem Biol 2015

Wilson et al. New Perspectives, Opportunities, and Challenges in Exploring the Human Protein Kinome Cancer Res. 2018, 78, 15-29

Wu P. et al. FDA-approved small-molecule kinase inhibitors. Trends Pharmacol Sci. 2015, 36,422-39.

Wu et al. Small molecule kinase inhibitors: an analysis of FDA-approved drugs, Drug Discovery Today 2016, 21, 5-10

Bio: Dr. May C. MORRIS, obtained her PhD in Biology and Health Sciences at the University of Montpellier in 1997 and completed her postdoctoral training at the Scripps Research Institute, La Jolla, USA. In 2000, she was hired by the CNRS to join the Centre of Research on Macromolecular Biochemistry in Montpellier. In 2005, she established her own research group and was promoted CNRS Research Director in 2000. In 2014 she moved to the Institute of Biomolecules Max Mousseron, where she currently heads the “Kinase Biosensors and Inhibitors” group within the team of Cellular Pharmacology. She has a solid scientific background on cell cycle kinases and cancer. She has established experience in peptide and protein biochemistry, in fluorescence spectroscopy to study protein/protein interactions, in cell biology and fluorescence imaging technologies, in peptide-based delivery and targeted therapeutic strategies. She has significant expertise in the design of fluorescent peptide biosensors to monitor protein kinase activity and in development of conformational protein biosensors implemented to screen for allosteric inhibitors.

She was awarded a CNRS Bronze Medal in 2006, a « Chercheuse d’Avenir » (Scientist of the Future) Award from the Region Languedoc Roussillon in 2009, MATWIN Oncology Prize in 2019 and Knight of the Legion of Honor in 2020. Her biosensor technology was recognized and certified by the Key Initiative “Biomarkers and Therapy” of Montpellier University of Excellence. She holds a solid publication record (>80 publications, Total citations 6214, H-factor 34 ISI Web of Science) and 8 PCT patent applications, edited a volume on Fluorescent Biosensors (Elsevier Press) in 2013 and a special issue on Fluorescent Biosensors in Biotechnology Journal in 2014. She has coordinated over 35 grants since 2000, including several multi-inter-disciplinary consortia. Editorial Board Member of ChemBioChem since 2010, Frontiers in Chemistry since 2015, Diagnostics since 2020. Member of the French Fluorescent Biosensor Workgroup of IMABIO GDR since 2011. In 2017 she coordinated and launched the regional initiative BIOCCI « Biosensors in Occitanie ».

Active member of Femmes & Sciences (French Women & Science) Association since 2008, she established the first Mentoring Programme for Female Doctoral students at the University of Montpellier, which became fully recognized in 2015, and coordinates the mentoring workgroup of Femmes & Sciences, contributing to disseminate the mentoring programme throughout French Universities, and becoming a member of eument-net network. In 2021, she was elected board member of EPWS (European Platform for Women in Science).