Reflex Biomarker Testing in NSCLC: Enabling Molecular-Guided Treatment from the Point of Diagnosis

When lung cancer is suspected based on imaging findings, a tissue sample is collected through a biopsy procedure, typically performed by an interventional pulmonologist, radiologist, or thoracic surgeon, depending on the location of the tumor. Using techniques such as bronchoscopy, CT-guided needle biopsy, or surgical biopsy, these procedures are invasive, can be challenging to perform, and often yield only limited amounts of tissue, particularly when tumors are located deeply within the lung.

The biopsy tissue is needed to support multiple analyses, including histology to establish the diagnosis and tumor type, and molecular testing to detect biomarkers that guide treatment planning. Because the amount of tissue obtained is often limited, the available sample may be exhausted during diagnostic workup, particularly if molecular analysis is delayed. In such cases, a repeat biopsy may be required, which is not always feasible due to the invasive nature of the procedure and because it may carry additional risk. This is a core challenge that reflex biomarker testing is designed to address.

Reflex biomarker testing began to emerge in the early 2010s in response to the rapid expansion of targeted therapies in non-small cell lung cancer (NSCLC), and refers to the automatic initiation of molecular testing by the pathology lab as soon as diagnosis of NSCLC is established, using the original biopsy specimen[1]. After the biopsy is performed, the tissue is sent to the pathology lab, where it is processed and examined to confirm diagnosis. In the traditional pathway, molecular testing is initiated only after the treating oncologist reviews the pathology report and requests specific biomarker analyses. This step introduces additional delays and may require retrieving stored tissue, by which point the available tissue may be limited.

By contrast, reflex biomarker testing integrates molecular analysis directly into the diagnostic workflow. Once the laboratory pathologist establishes a diagnosis of NSCLC, tissue can be allocated appropriately and molecular testing is initiated automatically, according to predefined protocols, without waiting for a separate request from the treating oncologist. This approach reduces the risk that the sample will be exhausted before molecular analysis is completed and helps ensure that comprehensive biomarker results are available when treatment decisions are made.

Through reflex biomarker testing, NSCLC biopsy samples are analyzed for key predictive biomarkers including EGFR mutations, ALK rearrangements, ROS1 fusions, BRAF, MET, RET, NTRK, KRAS, and PD-L1 expression, typically using next-generation sequencing (NGS) and immunohistochemistry (IHC)[2]. Performing these analyses at the time of diagnosis ensures that the molecular profile of the tumor is available when treatment decisions are made. This contrasts with the traditional sequential approach, in which biomarker tests are ordered individually after diagnosis, introducing additional turnaround time and potentially delaying the initiation of the most appropriate therapy[3].

Earlier identification of actionable alterations enables timely use of targeted therapies, which have been shown to improve clinical outcomes compared with non-selected treatment approaches. For example, EGFR mutations, present in approximately 15-20% of lung adenocarcinomas, can be treated with EGFR tyrosine kinase inhibitors (EGFR-TKIs) or a combination of EGFR-TKIs and chemotherapy, which have been shown to be superior to standard chemotherapy as first-line treatment in this population[4]. Comprehensive NGS approaches, particularly those incorporating RNA-based analysis, can further improve detection of actionable alterations by identifying gene fusions that may be missed by DNA-only methods and has been shown to increase the number of patients identified as eligible for targeted therapies[3].

As targeted therapies continue to expand, comprehensive molecular profiling at diagnosis has become an essential component of NSCLC care. Reflex testing ensures that all relevant biomarkers are assessed using the original biopsy specimen, supporting equitable access to guideline-recommended therapies and reducing the risk that testing is incomplete or delayed[5]. More fundamentally, reflex biomarker testing reflects both the evolution of lung cancer treatment and a recognition that the timing of diagnostic steps is clinically consequential. As the number of actionable biomarkers has grown, timely molecular profiling has become critical to selecting first-line therapy. Reflex testing represents an adaptation of diagnostic workflows to this new reality, ensuring that molecular information is available when it is needed to guide treatment. By embedding molecular profiling directly into the diagnostic pathway, reflex testing enables more efficient use of limited tissue and supports the delivery of truly molecular-guided care.

References:

1. Gosney, J. R., Paz-Ares, L., Jänne, P., Kerr, K. M., Leighl, N. B., Lozano, M. D., … & Peters, S. (2023). Pathologist-initiated reflex testing for biomarkers in non-small-cell lung cancer: expert consensus on the rationale and considerations for implementation. ESMO open, 8 (4), 101587. doi: https://doi.org/10.1016/j.esmoop.2023.101587

2. Toth, L. J., Mokanszki, A., & Mehes, G. (2024). The rapidly changing field of predictive biomarkers of non-small cell lung cancer. Pathology and Oncology Research, 30, 1611733. doi: https://doi.org/10.3389/pore.2024.1611733

3. Zacharias, M., Absenger, G., Kashofer, K., Wurm, R., Lindenmann, J., Terbuch, A., … & Brcic, L. (2021). Reflex testing in non-small cell lung carcinoma using DNA-and RNA-based next-generation sequencing—a single-center experience. Translational Lung Cancer Research, 10 (11), 4221. doi: https://doi.org/10.21037/tlcr-21-570

4. https://pmc.ncbi.nlm.nih.gov/articles/PMC6280904/

5. Smith, B. F., Hampel, K. J., & Sidiropoulos, N. (2024). Benefits of Implementing Reflex Genomic Analysis for Nonsmall Cell Lung Cancer. The Journal of Applied Laboratory Medicine, 9 (1), 28-40. doi: https://doi.org/10.1093/jalm/jfad104

Jacek Antas

Chief Executive Officer

Jacek Antas is a shareholder of the Company, has spent more than 25 years in the financial services industry holding various positions in sales and consulting.

Mr. Antas obtained a master’s degree from the Warsaw School of Economics and has served as a board member of various
companies throughout his career.

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James B. Murphy

Chief Financial Officer

Mr. Murphy is a certified public accountant with over thirty years of experience in finance and operations management. He is currently a consultant with Danforth Advisors LLC (“Danforth”), a leading provider of outsourced strategic and operational specialists across functions in the life sciences industry. While at Danforth, Mr. Murphy has served over fifteen private and publicly held life sciences companies as CFO and CFO Advisor, helping them secure over USD 0.5 billion in financing and successfully execute pivotal asset transactions. Mr. Murphy functions as a consultant to Helix pursuant to a consulting agreement between the Company and Danforth.

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Thomas Mehrling

Medical Adviser

Thomas Mehrling (PhD in Pharmacology and MD) has over 20 years’ experience in multinational Pharma companies developing novel oncology compounds from preclinical research through to registration. Prior to entering the industry, he spent 13 years as an MD at the University Hospital in Frankfurt, working on preclinical and translational projects. He served as Director of European Oncology at Mundipharma International (2003–2013), building the company’s first European oncology business from the ground up out of Cambridge, UK, and completing the clinical development, registration and launch of two major products in Europe, DepoCyte® and Levact® (Ribomustin® and Treanda®). In 2013, he led the establishment of the Mundipharma Group’s start-up, Mundipharma EDO, developing anti-cancer therapeutics for solid tumours out of Basel, Switzerland.

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Kim Gaspar

Director Quality Assurance

Kim is the Director of Quality Assurance at Helix BioPharma Corp. An experienced quality assurance professional with expertise in Canadian, US, and EU regulations, she has been involved in all aspects of Phase I/II biopharmaceutical product development over the years, including regulatory submissions, QC laboratory compliance, tech transfer and third-party oversight of CMC activities, clinical QA, and bioanalytical data analysis. Kim joined Helix in 2000, transitioning into QA in 2003. She holds a B.Sc in Biochemistry and a Ph.D in Veterinary Physiological Sciences, both from the University of Saskatchewan.

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Brenda Lee

Director Clinical Operations

Brenda is the Clinical Operations Director at Helix Biopharma Corp. A clinical research operations professional with 25 years of experience managing clinical trials, ranging from early Phase I to late Phase IIIb/IV studies, she brings experience in clinical study protocol writing and development, trial start-up and vendor management, and a proven track record in planning and managing clinical trials to quality standards, timelines and budget. Brenda joined Helix Biopharma Corp. in 2018, working to advance the clinical program of L-DOS47. She holds B.Sc and M.Sc. degrees from the University of Toronto, specializing in Nutritional Sciences and Human Biology.

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Jerzy Leszczynski

Director

Jerzy Leszczynski is a shareholder of the Company, has spent more than 35 years developing businesses and has served in the capacity of board member of various real estate development companies. Mr. Leszczynski obtained his Master of Science in Chemistry from the Warsaw Institute of Technology.

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Janusz Grabski

Director, Chair of Audit Committee

Janusz (John) Grabski is a lawyer specialized in corporate and real estate law with over twenty years of experience.

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Malgorzata Laube

Director

Malgorzata Laube has over 19 years of experience in nuclear medicine. In her last role with Alberta Health Services, she was the Department Supervisor, Nuclear Medicine at Royal Alexandra Hospital. Ms. Laube obtained a MSc degree in Environmental Engineering from the Warsaw University of Technology and is based in Edmonton, Alberta, Canada.

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Jacek Antas

Chairman of the Board

Jacek Antas is a shareholder of the Company, has spent more than 25 years in the financial services industry holding various positions in sales and consulting.

Mr. Antas obtained a master’s degree from the Warsaw School of Economics and has served as a board member of various
companies throughout his career.

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Jonathan Davis

Advisor, ADC Discovery

Jonathan Davis received his Ph.D. from University of California, San Francisco, where he studied protein structure and function using NMR. After a post-doc at Harvard Medical School exploring RNA selection and structure in the labs of Jack Szostak and Gerhard Wagner, he went to work at EMD Serono, where his work involved improving antibody-based therapeutics, inventing a platform technology for generating heterodimeric Fcs as a basis for multifunctional molecules, and developing a novel scaffold based on an artificially-designed protein from David Baker’s lab. In 2008 he took a job at Bristol-Myers Squibb in Waltham/Cambridge MA, working on antibody discovery and platform development in a wide range of therapeutic areas, with a particular focus on multispecific therapeutics. He moved to Madison, WI in 2019 to take on the role of VP of Innovation and Strategy at Invenra, a biotech focused on bispecific antibodies, and where he is currently head of the Scientific Advisory Board. In early 2024 he left the corporate world to found Creative Antibodies, a consulting firm that helps guide companies to successful antibody discovery and development projects, from mAbs to multispecifics, ADCs, and other formats. Outside of science, Jonathan is a conservatory trained cellist, plays numerous other instruments, and founded the UCSF Orchestra (now Symphony Parnassus) in San Francisco, where he was Music Director for six years.

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Davide Guggi

Advisor, CMC

Davide graduated as a pharmacist and received his PhD in Pharmaceutical Technology and Biotechnology from the University of Vienna. He has over 20 years of experience in the pharmaceutical industry, principally in the field of oncology. At the beginning of his career, Davide led oncology business units and commercial departments at Mundipharma and Gilead across Austria and Eastern Europe. Since over 10 years he has been working as a CMC expert, covering operational and regulatory CMC functions on behalf of over 20 different small- and medium-sized biotech companies across the world. He has served as CMC Director and CSO/CTO for several years, developing both small molecules and biologics (mABs, Fab, ADCs and Radio-immuno-conjugates) from early discovery to NDA/BLA in the US, EU and Canada, with a focus on First-in-Human and Phase I/II studies in oncology indications.

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