NewsRelease

AMini-Review Describes the Analytical Aspects of Liquidchromatography – Mass spectrometry (LC-MS) Methods Aimed atQuantification of Stroma Liquid Biopsy™ Proteins in Body Fluids

MONMOUTHJUNCTION, NJ, January9, 2023 -- Biotech Support Group (BSG) reports on joint publicationwith authors from research collaborators at LeidenUniversity Medical Center,describingthe developmental and analytical aspects ofTier 2 LC-MRM-MS protein quantification methods for liquid biopsies,with a special focus on BSG’s Stroma Liquid Biopsy™ proteinpanel. Thecitation is:


N.Diederiks, C.J. Ravensbergen, M. Treep, M. van Wezel, M. Kuruc, L.Renee Ruhaak, R.A.E. Tollenaar, C.M. Cobbaert, Y.E.M. van der Burgt,W.E. Mesker, Developmentof Tier 2 LC-MRM-MS protein quantification methods for liquidbiopsies,Journal of Mass Spectrometry and Advances in the Clinical Lab (2022),https://doi.org/10.1016/j.jmsacl.2022.12.007

Inthe pursuit of personalized diagnostics and tailored treatments,quantitative protein tests contribute to a more precise definition ofhealth and disease, driven by unmet clinical needs. Internationallystandardized TNM classifications of malignant tumors are beneficialfor diagnosis, however treatment outcome and survival of cancerpatients is poorly predicted. To this end, the importance of thetumor microenvironment (TME) has endorsed the introduction of thetumor-stroma ratio as a prognostic parameter in solid primary tumortypes, determined via routine diagnostic pathology slides. However,with the development of liquid chromatography – mass spectrometry(LC-MS) methods we aim at quantification of tumor-stroma specificproteins in body fluids, as the TME comprises a dynamic ecosystem ofcellular and subcellular interactions that surrounds malignant cells,essential to tumor progression. The article states “Despitesignificant advances in malignant cell-derived biomarkers, such ascirculating tumor cells (CTCs) and circulating cell-free tumor DNA(ctDNA), the TME has remained surprisingly underappreciated incurrent liquid biopsy research. To address TME profiling in liquidbiopsy, the Stroma Liquid Biopsy™ (SLB) panel was developed as anexperimental stroma-oriented proteomics alternative to theconventional genomic liquid biopsy biomarkers in oncology”.

“TheSLB panel comprises a set of key proteins in interconnected stromalpathways (e.g. coagulation, acute phase inflammation) and is believedto capture a deranged systemic response to the presence of cancer ina plasma proteome blueprint. Remarkably, we recently demonstratedthat the representing genes of the SLB protein panel composeexpression signatures that are expressed in malignant cells and thenon-malignant cells within the TME in colon cancer. Plasma proteinlevels of the SLB panel may, therefore, reflect histologicintratumoral stromal content and provide valuable prognosticinformation in liquid biopsies.”. A Tier 2 liquid chromatographymultiple reaction monitoring mass spectrometry (LC-MRM-MS) assay, toidentify and quantify proteins from the SLB panel requireswell-defined selectivity, repeatability, sensitivity, andreproducibility and is the optimal choice for analytical performanceof potential biomarkers. Inthis mini-review the analytical aspect of this developmentaltrajectory is further detailed.

Aimsand key objectives of the BSG – Leiden Research Collaboration

Tumorsare more than just a mass of proliferating cells and therefore cancerprogression is influenced by the multiple normal non-cancerous celltypes and networks of proteins dynamically interacting in activetumorigenesis. These are not simply passive bystanders andconsequently reflect the tumor-associated microenvironments calledthe stroma. Percolating through the tumor mass and introduced intothe general circulation, stromal conditioning of tumors can thus bemonitored by blood tests. Accounting for and characterizing thestromal influence on disease progression and response to therapieswill become of paramount importance in the management of cancerpatients in the future.

Basedon stained surgically removed primary tissue sections, LUMC hasdeveloped a tumor-stroma ratio (TSR) reported to be a strong,independent prognostic parameter linking tumors with high stromalcontent to poor prognosis inmultiple solid primary tumor types. In addition to patient prognosis,the TSR was reported to predict response to neoadjuvant therapy inbreast andgastrointestinalcancers. Lastly, the combined assessment of the TSR and histologictumor immune cell infiltrate was recently suggested as a novel toolin predicting response to checkpoint inhibitor therapy in coloncancer. These reports support the notion of the TME as a valuableaddition to current diagnostics in clinical oncology. Given the lackof TME parameters in cancer staging and grading classifications, theTSR is currently subjected to prospective validation in colon cancerin the international UNITED trial.

Thepatent pending panel of BSG’s Stroma Liquid Biopsy™ proteomicbiomarkers, in part reflects the tumor-associated microenvironmentsintroduced into the bloodstream, and to which can be monitored byblood tests. In this project, we aim to characterize and quantify theStroma Liquid Biopsy™ biomarkers from LUMC collected patient sera,so as to correlate to the LUMC developed Tumor-Stroma Ratio scoringmethods. Once achieved, we will have gained invaluable informationcentral to understanding how individuals uniquely adapt to thepresence of cancer anywhere in the body, how individuals can bestratified towards the best therapeutic options, and how individualsuniquely respond to medical intervention.

ToLearn More About Tumour-Stroma Ratio, visit:http://www.watchstroma.com

ToLearn More About Stroma Liquid Biopsy™, download whitepaper at:

StromaLiquidBiopsyWhitepaper02162022.pdf(biotechsupportgroup.com)

AboutBiotech Support Group LLC

Convergingwith cultural and technological disruptions forthcoming inhealthcare, Biotech Support Group develops methods for cost effectiveand efficient sample prep essential for these expanding markets.Following a tiered business strategy, the company continues itsgrowth in the consumable research products area supporting therapidly expanding installation of LC-MS instrument and computationalinfrastructure. For this market, key products include: AlbuVoid™and AlbuSorb™ for albumin depletion, Cleanascite™ for lipidadsorption, and HemogloBind™ and HemoVoid™ for hemoglobinremoval. From these innovations, the company has acquiredknowledgebase and biomarker intellectual property assets that supportdiscoveries of protein markers from blood, with special emphasis onearly detection and personalized medical decisions for cancerpatients. For more information, gotohttps://www.biotechsupportgroup.com/Default.asp

Forbusiness development at BSG,

Contact: MatthewKuruc 732-274-2866,[email protected]

AboutLeiden University Medical Center (LUMC)

Asthe world changes and its population ages,thenumber of patients with chronic diseases and disorders is expanding,while medical costs increase. Therefore, the health of the globalpopulation is one of the biggest challenges of our time. LeidenUniversity Medical Center believes that this challenge asks for aclear vision and mission. At the LUMC, students are trained inlifelong innovation and learning. Researchers let themselves beinspired and test their findings directly in practice. Patients cancount on being helped according to the state-of-the-art in science.The LUMC believes in curing but also in prevention. By collaboratingwith companies and organizations inside and outside the region, theLUMC makes innovations and new applications actually possible. Thusthe LUMC serves as an innovator for improving healthcare science andconsequently people’s health. For more information, gotohttps://www.lumc.nl

Keywords: StromaLiquid Biopsy™, Tier2 LC-MRM-MS, TumorMicroenvironment (TME), Tumor-StromaRatio (TSR), Cancer patientprognosis, predicting response to checkpoint inhibitor therapy