Clinical validation of Trop-2 as a serum biomarker for monitoring of disease-course in patients affected by breast and colon cancer

Project Information

ThruBlood

Grant agreement ID: 719856

DOI

10.3030/719856

Project closed

EC signature date 22 February 2016

Start date 1 March 2016

End date 31 August 2016

Funded under

• SOCIETAL CHALLENGES - Health, demographic change and well-being

Total cost

€ 71 429,00

EU contribution

€ 50 000,00

Coordinated by

ONCOXX BIOTECH SRL
Italy

Periodic Reporting for period 1 - ThruBlood (Clinical validation of Trop-2 as a serum biomarker for monitoring of disease-course in patients affected by breast and colon cancer)

Reporting period: 2016-03-01 to 2016-08-31

Summary of the context and overall objectives of the project

Cancer is the leading cause of death globally.
Current follow-up approaches adopted for the screening of patients in the post-treatment phase of breast/colorectal cancer rely on technologies that are affected by strong limitations in terms of cost-effectiveness (i.e. imaging technologies) and performance (e.g. serum biomarkers like HER2 for breast, CA-125 for ovary and CEA for colon).
Moreover, the drivers of disease recurrence and metastases are markedly heterogeneous and often depend on features of primary cancer such as biological aggressiveness, response to therapy and prognostic features. As a result, the set-up of effective therapeutic strategies for individual patients is remarkably challenging.
This reduces the efficacy of administered therapies, is at risk of considerably reducing the quality of life of the patient and poses a heavy management and financial burden on the health system.
To achieve the goal of effective choice among effective therapies, novel cancer biomarkers with high specificity and sensitivity are urgently needed.
Such biomarkers will allow a timely and accurate monitoring of the disease course of patients bearing, e.g. breast or colorectal cancer that have been diagnosed and treated.
After over 10 years of research, the staff of Oncoxx Biotech, a Cancer Biotech Company, has discovered in cancer patients and pre-clinical models that Trop-2 is the only gene consistently expressed at high levels in the majority of cancer cells that generate metastatic tumours in vivo. With this information at hand, we started our studies to determine whether Trop-2 also is a circulating, prognostic marker of tumors. We found that this protein is consistently released in the bloodstream, when a subject is developing a relapse from most of the analyzed cancers. This is opening a completely new pathway for introducing a low-cost and widely available serum biomarker for monitoring the disease progression.
Within this R&D we have created three monoclonal proprietary antibodies anti-Trop-2, as a possible quasi universal cancer target.
The antibodies differentially inhibit growth of (i) primary tumors and (ii) metastatic cells in pre-clinical models and synergize in vivo.
The top two biomarker’s features are high specificity (100%) and high sensitivity (80%), that coupled with a low-cost, will enable the effective implementation of the novel high throughput and high accuracy assay through the most common diagnostic procedures, like the ELISA test.
This is a sound starting point, which positions Trop-2 beyond the biomarkers currently adopted, as it will allow: (i) the early detection of patients with aggressive tumours, (ii) the assessment of the tumour burden at any given stage of the disease, (iii) to monitor tumour-bearing patients to draft a relapse-risk profile. Our ambition is to revolutionize the way disease-course monitoring and treatment are conceived, allowing clinical laboratories to avail a reliable diagnostic tool capable to accurately identify those patients at high risk of developing metastatic relapse.
The overall objective of the ThruBlood project will be the Clinical Validation of the relationship of circulating Trop-2 levels with the biological characteristics of the tumor, to allow the best available therapy and the best monitoring of disease course.
The key strategic objectives of the Thrublood project Phase 1 were:

- Strategic Objective 1: design of the ThruBlood assay for the measurement of Trop-2 level in blood serum of cancer patients at surgery, 1 month after surgery and at 6 months intervals afterward. Trop-2 detection will be performed by a newly developed, state-of-the-art ELISA assay. Detection and levels of circulating Trop-2 will be correlated with clinical and pathological status. Five leading medical institutions will be involved from Italy, Germany, France and Croatia, to achieve high enrolment capacity, medical impact and ultimate success of the project

- Strategic Objective 2: Study of the Assay Validation: an innovative ELISA assay will be developed, with high specificity and sensitivity, coupled with a low-cost, high-throughput assay for effective implementation within a diagnostic procedure.

- Strategic Objective 3: Laboratory Validation of the the ELISA-ThruBlood assay. To validate procedures for measuring circulating Trop-2 in clinical settings in terms of (i) accuracy, (ii) reliability, (iii) predictive power, (iv) speed, (v) cost efficiency, to pave the way for extensive, routine use in cancer centers worldwide.

- Strategic Objective 4: design of the Clinical Validation trial: (a) To validate Trop-2 as a disease history predictor, through systematic measurement of Trop-2 in the serum, across large groups of breast, ovarian and colon cancer patients, at diagnosis and after surgery.

- Strategic Objective 5: analysis of the business potential of the ThruBlood assay in the market of cancer biomarkers.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The actions developed under the SME Instrument Phase 1 project were focused on the activities required for the design and implementation of the novel Cancer Biomarker Assay, on its Clinical Validation, to be possibly carried out under SME Instrument Phase 2 project, and on market research and planning of market entry, to allow to utilize the results achieved in the project for effective use of the Cancer Biomarker Assay in clinical settings.
In particular, we have carried out activities aimed at the fulfillment of the following strategic objectives:

Task 1 - Planning for clinical Validation in the Phase 2 project.
The objective was the identification of the clinical needs in the international scenario – biomarker validation for patient monitoring, early diagnosis and therapy planning for breast, ovary and colon cancer.
The work performed allowed us to to define the application scenario for the implementation of the ThruBlood biomarker within three target cancers (Breast, Colorectal and Ovarian). In this respect, we have carried out an extensive analysis of the current “follow-up” procedures adopted in Europe for monitoring patients treated for breast and colorectal cancer, through both a literature survey and direct interactions with European and International leading organisations. As a result of this study, we found that there is a common lack of specificity in the approaches currently adopted for the follow up of breast, colorectal and ovarian cancers, which entails difficulty to individuate patients at high risk of locoregional relapses.
This suggests a potential business case in all three target applications.

Task 2 - Design and validation of high-throughput systems for the assessment of circulating Trop-2 levels.
We started from the analysis of the proprietary ICONA system as platform for the development of ThruBlood assay. However, the first tests carried out with the assay suggest that the system still requires major engineering and testing activities to ensure performances in line with the requirements of a clinical trial, thus suggesting us that a more robust platform should be used. For this reason, we have analysed another platform suitable for the implementation of the Trop2 biomarker that is ELISA (Enzyme-Linked ImmunoSorbent Assay), a leading solution for high throughput screening.
In addition, to move from the current TRL (around 6) of the ThruBlood biomarker to its introduction into the clinical practices for follow-up of cancer patients and commercialization on the market (TRL 8-9), we decided that it was strategic to find a suitable industrial partner that could join the project, to support us in the development of the ThruBlood assay, thus facilitating its introduction in the market of In Vitro Diagnostics.
After a broad search at European level relating to companies working on the development and commercialisation of cancer biomarkers, the German company AJInnuscreen resulted as the most interesting, since they have both the technological know-how at industrial scale and the business potential adequate to support the development and commercialization of the new product. Thanks to the cooperation with AJInnuscreen, we were able to define the lay-out of the ThruBlood assay and the main activities to be carried out for the successful commercialization of the proposed biomarker.

Task 3 - Design of the Clinical Validation Study of the ThruBlood biomarker.
The main objectives of the clinical validation of the proposed biomarker are: (i) the early detection of patients with aggressive tumors, (ii) the assessment of the tumor burden at any given stage of the disease, (iii) to monitor tumor-bearing patients to draft a relapse-risk profile.
A screening of the existing oncological clinical centers across Europe has been carried out, in order to find clinical centers at the forefront of the research on the ThruBlood target diseases. In addition, we have involved into project the Italian CRO Consorzio Nazionale per la Ricerca in Medicina (namely CIRM), which is a nonprofit organization dedicated to the promotion of the medical research in Italy and full member of the ECRIN (European Clinical Research Infrastructure Network) Network.
Oncoxx will collaborate with AJInnuscreen to develop and validate the most suitable blood serum test protocol to be carried out in clinical settings. CIRM will cooperate with Oncoxx and AJ Innuscreen to validate such blood serum assay in clinical settings. CIRM will also provide all required indication as to the clinical requirements, clinical trials protocols and related certifications for the Trop-2 ELISA assay. The clinical validation was conceived to last 30 months, by considering June 2017 as starting date in case Phase 2 project granted, and it will comprise the following phases:
• Validation of Trop-2 as a disease history predictor, through systematic measurement of Trop-2 in the serum, across large case series of breast, ovary and colorectal cancer patients, at diagnosis and periodically over time after surgery.
• The validation will be performed at distinct pathological stages of breast, ovary and colorectal tumors, in order to obtain the most complete trials outcomes. Several aspects, such as age of the patients, and the metastatic disease progression path, previous therapy, performance status and other pathological indicators, will be considered.
We have prepared a clinical protocol in cooperation with CIRM containing the main features of the clinical validation trial, to be submitted for approval to local Ethical Commitees of clinical centers involved in the multicentric trial.
We have designed the operational plan by allocating WP and responsibilities to all the partners, along with the estimation of the project through the definition of a GANTT chart.

Task 4 - Business Opportunity.
We have studied the In Vitro Diagnostics (IVD) market, that is the target market of the Thrublood device, the size, the current grow trends and the main drivers that influence its expansion.
We assessed the value proposition delivered by the ThruBlood assay by analyzing the business cases related to the three cancer typologies we planned to validate during the clinical trial.
In this respect, we analysed key parameters such as the reimbursement policy of the regional health system, incidence of the specific cancer, and costs due to the adopted follow-up procedures.
We also redesigned the business model of Oncoxx Biotech and we made a projection of the future business model pursued after the Phase 2 project completion that will derive from the agreement with AJInnuscreen company.

The main outcomes of such actions are summarized as follows:

1. We have designed and finalised the project work plan whereby AJInnuscreen, Berlin, Germany will participate in the development of the final ThruBlood assay.
2. We have defined the final objectives of the Clinical Validation to be possibly carried out under SME Instrument Phase 2 project, taking into account both the starting considerations and the additional expertise and background coming from the participants involved in the WP.
3. We have designed and finalised the project work plan for the clinical validation and the allocation of responsibilities amongst the participants, which will be CIRM, Milano, Italy for the organisation and management of the clinical validation.
4. We have analysed the business opportunity for the ThruBlood product, given the current unmet needs and the market drivers that are responsible for the cancer biomarker market growth. A first agreement has been reached with AJInnuscreen, Berlin, Germany for carrying out a Phase 3 of the project for market entry and commercialisation of the final ThruBlood assay.
As a key example, the global market for Predictive Breast Cancer diagnostic and drug technologies was valued at $23.4 billion by a recent report, twice as much as that of the drug market, and is expected to rise at a compound annual growth rate (CAGR) of 2.5%, to reach $24 billion by the end of 2016. Perspectives are to have the potential to acquire a significant fraction of this market in regard to cancer diagnostics.

Output publications and patents
Project activities have been based on the scientific leadership of the Oncoxx team in the field of Trop-2, as a determinant of cancer growth, metastatic spreading and tumour patient outcome. Most important findings have been reported in prestigeous international, peer-reviewed journals, including seven publications in 2016. Inventive height and Intellectual Property protection activities have led to ten patents, six of which PCTs, three of which granted in the European Community, in the US and other major countries.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The performance of procedures for early detection of relapses and metastases required being pushed beyond their current limitations. Therefore, novel biomarkers with high specificity and sensitivity are urgently needed to monitor the disease course of patients diagnosed and treated for breast, ovary or colon cancers, aimed at identifying those are at the highest risk of developing metastases and relapses.
The goals of our study have been to develop an assay for measuring serum circulating Trop-2, as a means to detect disease appearance or relapse in cancer patients. Main biomarker’s features that have been developed have been high specificity and high sensitivity, coupled with a low-cost, high-throughput assay able to measure the biomarker in large patient's groups.
A first study was performed on patients with breast cancer, or with benign adenomas and on healthy donors. Our unprecedented results showed that Trop-2 is released in the bloodstream and that all higher-than-baseline levels of Trop-2 identify breast cancers with a specificity of 100% and sensitivity of 80%.
A second study was performed on 105 patients with colorectal cancer and on healthy donors and demonstrated that the higher-than-baseline levels of Trop-2 identify colon cancers with a specificity of 100% and a sensitivity of 74%.
A third study has been carried out to extend such findings to 'other cancer' and led to obtaining corresponding findings in ovarian cancer patients.
Notably, these findings have been achieved through easy-to-implement laboratory procedures, thus eliminating the need for additional transfer activities to specific clinical laboratory settings.
The activities that have been performed will seamlessly join the next phase of the study, i.e. the clinical validation of the Cancer Biomarker Assay. This will allow to transforming the Trop-2 cancer biomarker from valuable candidate to established technology for enabling oncologists to tailor treatment for individual patients, through a precision medicine strategy.
The expected impact of developing a novel, non-invasive, circulating cancer biomarker assay with the performance outlined above is very high.
Post-operative relapses and metastases are the leading cause of cancer-associated mortality. There cannot be a single strategy valid for a heterogeneous population, and the future will rely on technologies able to define individual risk assessment, to personalize cancer care to best formulate an efficient treatment plan for each individual patient.
Thrublood’s value proposition is to make available a new cancer serum biomarker, able to timely and accurately detect the metastatic relapse in breast and colon cancer patients. This will dramatically enhance the quality of care, as well as its cost-effectiveness, as measured through Quality Adjusted Life Years (QALY) and Disability-Adjusted Life Years (DALY). The adoption of this novel biomarker will improve the decision-making process and personalized patient treatment. Pharmaceutical companies involved in drug discovery, as well as Contract Research Organizations, will implement the biomarker for streamlining the development and validation of breast, ovarian or colon cancer therapies. Research centers involved in the research of the physio pathogenic mechanisms underlying cancer progression will additionally benefit from the insight provided by our cancer biomarker assay to accurately monitor disease biological characteristics and their impact on disease relapse.
The global market for Predictive Breast Cancer diagnostic and drug technologies was valued at $23.4 billion by a recent report, twice as much as that of the drug market, and is expected to rise at a compound annual growth rate (CAGR) of 2.5%, to reach $24 billion by the end of 2016. Perspectives are to have the potential to acquire a significant fraction of this market in regard to cancer diagnostics.

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