Please are you able to introduce yourself and tell us about your role at Owlstone Medical?

My name is Mariana Leal, and I’m a Lead Translational Scientist – Team Lead at Owlstone Medical. Before I joined Owlstone Medical, I worked for a very long time in academia, mainly within the oncology field after which in a drug discovery biotech. My work involved biomarker discovery and likewise understanding the mechanism of motion and resistance to several anticancer therapies using clinical samples and pre-clinical models.

I joined Owlstone because the thought of making a non-invasive test is a key motivator for any translational scientist working with difficult diseases resembling cancers. I used to be intrigued by the technology that Owlstone Medical was developing and the potential for addressing the challenge of an early cancer diagnosis as a part of a multidisciplinary team.

One among my key roles in Owlstone Medical involves bringing together the knowledge of biomarker discovery and cancer metabolism to develop probes that will probably be used as a part of minimally invasive or non-invasive tests to detect lung cancer.

Owlstone Medical is a world leader in breath biopsy for early detection and precision medicine. Are you able to tell us more about Owlstone Medical and the way the breath biopsies are a great tool for investigating biomarkers for the early detection of diseases?

Owlstone Medical goals to save lots of 100,000 lives and $1.5 billion in healthcare costs through Breath Biopsy. Our evaluation is targeted on volatile organic compounds (VOCs) present within the exhaled breath. These VOCs are the product of metabolic processes inside an individual’s cells and tissues, inside their microbiome, and from their response to environmental exposures. Since VOC production is linked on to metabolic activity, the compounds will be sampled quickly and noninvasively from breath, urine, or other bodily fluids. Changes in VOC levels happen on the very earliest stages of disease, so detection of the compounds can allow disease diagnosis before other physical symptoms turn into apparent.

Introducing Owlstone Medical – Breath Biopsy for Non-invasive BiomarkersPlay

One particular focus area for Owlstone Medical is surrounding lung cancer. Why did you select to particularly concentrate on this deadly disease, and the way can your biopsy technology be utilized on this research field?

Lung cancer is considered one of the leading causes of cancer-related death worldwide. A robust contributor to this fact arises from the issue in detecting lung cancer at an early stage. Widespread public screening programs for lung cancer targeting at-risk populations represent considered one of the best opportunities to enhance early detection.

Nonetheless, currently, the one suitable methods (e.g., CT scanning) are a limited resource that requires specialist capabilities and will not be easily accessible to nearly all of the population. A breath test for lung cancer represents a non-invasive, preferable approach for screening that may be cheaper, easy to make use of, and accessible than current options.

We’ve observed an increased variety of indeterminate pulmonary nodules (IPN) needing further management with the advance of the present screening programs mainly based on CT scanning. Although the concept of IPN varies amongst guidelines, a nodule is taken into account indeterminate if the etiology is just not apparent on a CT scan and is usually defined as non-calcified and solid components between 6-30mm in largest diameters.

When an IPN is identified on a CT scan, the probability of cancer is often estimated from a clinical prediction model and defined from the management guidelines. Several blood and imaging biomarkers have recently been developed to define the probability of IPNs; nevertheless, none has yet demonstrated improvements in patient outcomes. This highlights the necessity for complementary approaches (including a breath biopsy) to define whether a nodule within the lung is malignant.

Despite a growing body of evidence that demonstrates the potential for breath as a way to detect illnesses, there was limited progress in developing a breath test for cancer. In addition to VOCs, exogenous volatile organic compounds (eVOCs) probes have been explored by Owlstone Medical to focus on various diseases. Are you able to tell us more about eVOCs?How can they assist to tackle the limitation of the breath evaluation based on volatomics?  

eVOCs are compounds not normally found on breath at significant levels. Nonetheless, Owsltone Medical is using these exogenous compounds to explore how the body absorbs, metabolizes, or excretes these compounds, revealing dysregulated critical pathways in numerous diseases.

An example of an eVOC probe under investigation for the detection of chronic liver disease is using compounds which might be FDA “Generally Recognized as Secure “(GRAS) compounds, resembling some food additives. These eVOCs are administered orally, undergo metabolism within the body, and are excreted via breath. This approach is exciting not only because we will amplify the signal from key metabolic pathways but in addition since the eVOC probe must be secure to be a part of a breath test for diagnosis or monitoring.

The exogenous volatiles (resembling the food additives) sometimes will not be enough to focus on some disease-specific pathways; thus, we’re also developing probes with more complex structures. For instance, we conjugate the substrate of enzymes with a volatile tag (as presented on the AACR 2023 congress), and the discharge of this tag and its detection in breath is related to the presence of the goal enzyme that cleaved our probe.

Thus, eVOC probes enable the event of research tests for applications where naturally occurring VOCs alone could also be insufficient and make it possible to run smaller, more focused, and potentially faster clinical trials for us and our customers.

Image Credit: Owlstone Medical

At AACR, you might be presenting a research poster that investigates the breath-based detection of lung cancer using exogenous volatile organic compounds (eVOCs) in humans. Are you able to talk us through the way you conducted this research and your preliminary findings? 

At AACR, we presented the outcomes of our clinical trial Evolution, an eVOC proof-of-concept study. Within the Evolution study, we evaluate whether administering a probe compound, specific to tumour-associated extracellular β-glucuronidase, produces a novel EVOC on breath of a patient with lung cancer.

We developed D5-ethyl-βD-glucuronide as a hydrophilic substrate probe that, upon hydrolysis by β-glucuronidase, releases D5-ethanol as a novel volatile reporter. We confirmed the cleavage of our probe by the goal enzyme and, due to this fact, the detection of the deuterated labeled ethanol using our in vitro headspace system. It will be significant to focus on that the same probe (same mechanism) was previously tested in in vivo cancer mouse models (Langer et al. 2019). At Poitiers University, it was previously observed that cancer-bearing animals treated with a D5-ethyl-βD-glucuronide excrete high levels of D5-ethanol greater than healthy animals, and the degrees of D5-ethanol observed were proportional to the tumor volume.

To ascertain the concept, we accessed the utility of our goal probe by evaluating β-glucuronidase activity in samples from a lung cancer PDX model and the protein expression by immunohistochemistry using a big cohort of samples from human primary lung tumors, lymph node metastases, and otherwise normal lung tissue. Immunostaining of lung resection specimens showed elevated levels of extracellular β-glucuronidase in 88% of stage 1 non-small cell lung cancers, 83% of stage 2, and 95% of stage 3. For adjoining non-malignant tissue in lung cancer subjects and non-malignant lung tissue resection in controls, staining was restricted to intracellular sources, particularly inside macrophages.

D5-ethyl-βD-glucuronide was then administered to 43 individuals to ascertain safety and evaluate eVOC levels on breath using various breath collection and analytical approaches. Intravenous administration of the probe to subjects with lung cancer and controls revealed a superb safety profile. The cleavage product D5-ethanol could possibly be detected on breath in a subset of participants, and this showed a relationship with the amount of breath collected. This study demonstrates proof of mechanism for the in human cleavage of the volatile reporter molecule D5-ethanol from D5-ethyl-βD-glucuronide.

What are the subsequent phases of this study?

The outcomes from these proof-of-concept studies provide a promising foundation for a phase 2 dose-finding study designed to explore the diagnostic performance of this modern breath test approach for lung cancer. We’ll proceed our clinical study by testing a low probe concentration and optimizing one of the best time point for breath collection to optimize the signal-to-noise ratio of the breath test and with the aim to discriminate patients with and without lung cancer. It will be significant to focus on that, for the subsequent phase, we are going to use an optimized approach to breath collection and evaluation to extend the sensitivity to detect our goal VOC even further.

For this research poster, you partnered with leading institutions and organizations within the oncology space, including Cancer Research UK, the NHS, and the University of Cambridge. How vital is collaboration to your work at Owlstone Medical, and the way do strong relationships and partnerships speed up difficult research areas resembling cancer?

Working with others allows us to speed up the sphere of breath evaluation. We work closely with the important thing opinion leaders, ensuring we’re investigating clinically relevant populations to handle relevant biological questions and developing products that may impact the patient’s life positively, including within the oncology field, where we urgently need recent technology for early diagnosis.

Image Credit: Giovanni Cancemi/Shutterstock.com

What’s next for you and your work at Owlstone Medical within the oncology field? Are you involved in any exciting upcoming projects?

One among the exciting upcoming projects involving lung cancer diagnosis is the brand new research agreement with Bicycle Therapeutics plc. We’ll explore the event of antigen-targeted diagnostic probes that use bicyclic peptides as their targeting mechanism linked with Owlstone’s EVOC® probes. It is a proof-of-principle study for the broader opportunity by promoting the selective accumulation of the probe on the tumor for increased signal and enhanced specificity.

If successful, our work has the potential to support not only the diagnosis of the patient and speed up the work to construct a more complex panel for lung cancer screening. As well as, in clinical trials, our probes will be potentially used for patient stratification. Due to this fact, we expect to expand to companion diagnostic tests to discover responders/non-responders for therapy selection and to measure goal engagement throughout treatment.

Seeking to the longer term, you might be aiming to expand your pre-clinical system capabilities. How are you aiming to do that, and what do you hope to attain from this expansion?

Owlstone Medical is constructing a novel platform to find VOCs related to key metabolic pathways related to a disease but in addition a strong pipeline to find recent eVOC probes. We currently can evaluate liquid or air headspace VOC from in vitro or ex-vivo studies, and we’re also developing different methods to gather breath samples from mice. The important thing point is that we use a system to trap and/or analyze VOC in our pre-clinical studies just like the one we use for Breath Biopsy, highlighting the strong translational potential of VOC evaluation from pre-clinical to clinical phases.

Strategies for VOC evaluation in in vitro, ex-vivo, and in vivo animal studies will be targeted or untargeted. The untargeted approach involves measuring the entire spectrum of VOCs released within the headspace or animal breath within the presence or the absence of a disease or treatment of interest. The untargeted approach is right if the VOCs alterations are unknown. However, targeted approaches measure specific known compounds, generated either by cellular metabolism or by treatment with substances metabolized by metabolic pathways of interest, as our eVOC probes.

Analyzing VOCs produced by in vitro, ex-vivo, or in vivo, models can display mechanistic links between disease biology and biomarker candidates. These can subsequently be verified in clinical studies, sampling the identical VOCs on breath, providing vital evidence that would bring diagnostic breath tests into clinical practice.

With increased research efforts surrounding diseases, each respiratory and cardiovascular, we’re seeing scientific breakthroughs becoming more commonplace. What do you suspect the longer term of breath biopsy to seem like? Are there any particular developments you might be excited to see?

Metabolic, inflammatory, and infectious diseases have the potential to be diagnosed or monitored by breath evaluation. With sufficient biological understanding underpinning them, breath tests can due to this fact goal specific metabolic pathways. With further knowledge of those pathways, we will perform targeted evaluation using our platform, providing confident identification and quantification of a panel of VOCs in breath.

Using different techniques, Owlstone Medical is exploring biomarkers for various lung diseases in clinically relevant populations, including lung cancer, infection, and other inflammatory processes. I’m excited to see where we will pull all these pieces together to assist in the elucidation of IPN needing further management. As we will not be directly assessing the placement of a certain disease, as in an imaging test, we recognize that the longer term of a diagnostic breath biopsy test may require a panel of eVOC probes together with goal VOC evaluation to have more robust and reproducible tests.

Image Credit: SciePro/Shutterstock.com

Are you hopeful that with continued advancements surrounding breath biopsies, we are going to see earlier detection rates for lung cancer?

Yes! As discussed above, widespread public lung cancer screening programs mainly goal the population defined as high-risk and depend upon CT scanning. The success of those screening programs strongly will depend on the engagement of the goal population accessing the imaging sites, a widely known barrier. As well as, the screening programs for lung cancer mainly goal the population currently defined as high-risk, normally defined by age, smoking, and family history. Optimization of the population to be screened still must be developed. Due to this fact, breath biopsy may also help to beat the currently limitations in lung cancer diagnosis and screening.

In addition to offering your research services, several resources can be found in your site, including publications, posters, eBooks, and blogs. Why is offering a wide selection of resources vital to you?

Owlstone Medical’s vision is to be the leading source for biomarker discovery on breath, providing tools for researchers. Furthermore, we’re constructing a catalog of the validated and quantitated VOCs found on breath: the Breath Biopsy VOC Atlas, which incorporates insight and the scientific context of their identification. We wish to supply one of the best discovery solutions available to our customers and maximize the possibilities of finding biomarkers with the potential to be translated into clinically invaluable tests.

As well as, we host a Breath Biopsy Conference annually to extend the discussion and collaboration around Breath Biopsy and VOC identification.

Where can readers find more information?

Find more concerning the study presented at AACR 2023: https://www.owlstonemedical.com/media/uploads/files/2023-04_Evolution_poster_AACR_Update_FINAL.pdf

Find more about VOC evaluation here: https://www.owlstonemedical.com/resources/

Also follow Owlstone Medical news here: https://www.owlstonemedical.com/about/news/

More about opened positions here: https://www.owlstonemedical.com/about/careers/opportunities/

About Mariana Leal

Dr Mariana Ferreira Leal is a Lead Translational Scient at Owlstone Medical.

Initially of her profession, Dr Leal focused on the investigation of gastric cancer diagnostic and prognostic biomarkers. She worked with gastric cancer for 15 years and led projects before moving to Royal Marsden Hospital (RMH)/The Institute of Cancer Research (ICR) in London.

She was a postdoc fellow with  Dr Mitch Dowsett after which with the Dr Lesley-Ann Martin team with expertise in phenotypic and genotypic characterization of ER+ breast cancer clinical samples and in vivo and in vitro models for biomarker discovery and validation, coupled with data evaluation and interpretation. She investigated the mechanisms of response and resistance to several goal therapies and identified mutational and transcriptional markers that predict patients who may gain less profit from endocrine therapies alone. She generated disease linkage data to guide indication selection and possibly targeted patient populations. She led studies with clinical trial samples in addition to one longitudinal retrospective study involving patients treated on the RMH, during which she put together the cohort based on RMH databases.

This data brought her to work as a Principal Scientist in a drug discovery biotech specializing in the modulation of transcription aspects in breast cancer.

And now at Owlstone Medical, Dr Leal is a lead translational scientist focused on the invention and development of non-invasive tests for lung cancer diagnosis and/or screening based on Breath Biopsy. Her role involves the invention of recent enzymatic targets and de-risk exogenous volatile organic compound (eVOC) probes to deliver biomarkers for brand new clinical studies specializing in non-invasive lung cancer diagnosis. She designs and leads several in vivo and in vitro studies, including using a set of biological and chemical tools to modulate the enzymatic activity of our targets. She also supports clinical trials during their completion.

Dr Leal has published extensively, with a complete of 89 publications, beside book chapters and conference related documents. She is a member of varied skilled societies, including the eACR and ASCO.