MOLECULARLY IMPRINTED POLYMER-SENSOR OF URINARY MODIFIED NUCLEOSIDES FOR NON-INVASIVE THERAPY CONTROL OF CANCER – CancerSensor

This project aims to develop and validate a quantitative, non-invasive diagnosis tool to monitor the efficacy of chemotherapy in patients with colorectal cancer and to follow up these patients after surgical resection, even at a metastatic stage. An acoustic-biosensor based on molecularly imprinted polymers (MIP) was obtained. It includes advances which will take the sensitivity down to a low-concentration single-molecule level. Apart from detecting and identifying tumor markers in the urine samples, the effect of circadian chemotherapy on the amount of cancer markers, using the developed tools will be explored. In fact, the circadian timing system can modify 2- to 10-fold the tolerability of anticancer medications in experimental models and in cancer patients. This is to date an almost unexploited dimension for diagnosis information. By combining and supporting these novel optical methods with state-of-the-art affinity molecule MIP technology, tumor biomarkers, acoustic biosensor, will be exploited to extract a maximum amount of information out of very small amounts of sample material. Given the high incidence of colorectal cancer, this project has a very high relevance. In these conditions the utility of the development of rapid, sensible, easy-to-use and cost effective sensors for surveillance of pre- and post-operative CRC is evident and of huge interest. This approach could however deeply modify the manner of checking near each person the effectiveness of the chemotherapy treatment of colorectal cancer. At the end of this project, we will offers diagnostic products for colorectal cancer follow-up in the European market. It will have further the ability to detect and recognize the expression of tumor-associated biomarkers in patients with metastatic cancer.

Main data obtained :
? Sensitive and selective polymers of cancer biomarkers pseudouridine and N7-methylguanosine,
? Development of MIP solid phase extraction for extraction of nucleosides from urine,
? Development and validation of a SAW-MIP for polar nucleoside monophosphate,
? The proof-of-concept for a SAW-MIP detection of AMP from urine is successful,
? Development of ultrasensitive detection of urinary nucleosides by mass spectrometry,
? Approval of 3 clinical studies and sample collections from patient treated for colorectal cancer.
• Protocole C11-59 (chemotherapy) -> 16 patients enrolled
• Protocole C11-58 (metastasis resection) -> 7 patients enrolled
• Protocole C11-42 (circadian cycle) -> 29 patients enrolled

optimization of the current prototype
Application to othe urinary nucléosides biomarkers of other cancers (breast, lung, ...)

Data obtained from the various aspects of this transdisciplinary research were published as three PhD thesis, 6 articles in international peer-reviewed journals, 6 communications (mainly if international conferences) and 15 posters (mainly if international conferences). Most of this publications were co-signed with at least two teams from this consortium.

Colorectal cancer is the second cause of cancer-related deaths. Once colon cancer spreads beyond the colon, into the lymph nodes or into other areas of the body, it is then harder to treat. It is thus very important to can detect as earlier as possible a cancer, to treat it fast, to check for the efficacy of treatments and for any recurrence. The therapeutic drug monitoring of antineoplastic agents is at this time restricted by several factors, such as the lack of established therapeutic concentration ranges, a considerable inter- or intra-individual pharmacokinetic variability, a relationship linking toxicity to systemic exposure, and complicate pharmacodynamics of drug toxicity. To circumvent those limitations and evaluate the success of chemotherapy is to correlate a cancer treatment with the amount of tumour markers, a substance that can be found in the body when cancer is present. It has been reported that cancer patients excrete in their urine an abnormal increased amounts of modified nucleosides, which are formed at the post-transcriptional stage by chemical modification of normal nucleosides within the RNA. These modified nucleosides cannot be reutilized or further degraded, but they are excreted in the urine as intact molecules. The elevated levels of modified nucleosides in the urine samples have served as potential cancer biomarkers in many studies. Although different analytical techniques have been reported for determination of nucleosides levels, they are practically difficult to use as a routine tool for therapy control of colorectal cancer.

Goals of this proposal are to develop and validate a quantitative, non-invasive diagnostic tools to follow-up the chemotherapy of colorectal cancer and monitor the disease response to chemotherapeutics. A methodology will be developed making use of a combination of the probably most exciting recent advances in the field of acoustic-biosensor based on molecularly imprinted polymers (MIP). It includes advances which will take the sensitivity down to a low-concentration single-molecule level. Apart from detecting and identifying tumour markers in the urine samples, the effect of circadian chemotherapy on the amount of cancer markers, using the developed tools will be explored. This is to date an almost unexploited dimension of diagnostic information. By combining and supporting these novel optical methods with state-of-the-art affinity molecule MIP technology, tumour biomarkers, acoustic biosensor, will be exploited to extract a maximum amount of information out of very small amounts of sample material. Given the high incidence of colorectal cancer, this project has a very high relevance. Those tools will be used, within the frame of this project, to analyze more than 3000 urinary samples from patients under chemotherapy for colorectal cancer and to correlate the chemotherapy treatment with the circadian time of infusion and the circadian variations of clearance.

The need for development of rapid, sensible, easy-to-use and cost effective sensors for surveillance of pre- and post-operative CRC is evident and of huge interest. This approach could deeply modify the manner of checking near each person the effectiveness of the chemotherapy treatment of colorectal cancer. At the end of this project, we will offers diagnostic products for colorectal cancer follow-up in the European market. It will have the ability to detect and recognize the expression of tumour-associated biomarkers in patients with metastatic cancer.

To obtain the level and width of expertise required, we have thus formed a consortium involving highly recognized teams:

Organic and medicinal chemistry (Pr Agrofoglio – Orléans),
Biosensor technology (Pr Rebière – Bordeaux),
Bioanalysis (Dr Bénech - CEA),
Oncology and circadian biology (Dr Lévi – INSERM, Hôpital Paul Brousse - Villejuif),
Polymers and product development (Dr Vidal - MERCK Chimie – ESTAPOR)