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​ DxPG80 IVD ELISA Human circulating Progastrin (hPG80) Immunoassay​ The DxPG80 test is an ELISA (Enzyme-Linked Immunosorbent Assay) for the detection of hPG80 (circulating human progastrin). The test is intended for professional laboratory use only. ​ Please note that for greater clarity, we use the name hPG80, referring to extracellular Progastrin. The test is based on the principle of a sandwich ELISA to measure the concentration of hPG80 in plasma samples that have been collected in EDTA tubes. Briefly, a capture antibody specific for hPG80 is immobilised on the 96-well plate. The hPG80 present in the calibrators and samples added to the wells will bind to the immobilized capture antibody. The test plate includes calibrators which are used to estimate the level of hPG80 in EDTA plasma samples. The wells are washed and an anti-hPG80 detection antibody coupled to horseradish peroxidase (HRP) is added, resulting in an antibody-antigen-antibody complex. After a second wash, a substrate solution of 3,3',5,5'-Tetramethylbenzidine (TMB) is added to the wells, producing a blue colour directly proportional to the amount of hPG80 present in the initial sample. The Stop Solution changes the colour from blue to yellow, and the wells are read at 450 nm with a microplate reader. The assay is technically characterised in manual mode by a limit of detection (LoD, 1.0 pM) and a limit of quantification (LoQ, 3.3 pM). The LoD detects the presence of hPG80 and the LoQ gives the concentration at which the assay is quantitative. ​ Table 1 : Comparative description of hPG80 in Controls (healthy blood donors 18-40 years old at low risk of cancer) and Cancer patients (16 cancers tested, all combined)[1]. Reference range are given for a 2.5 (low range) and 97.5 (high range) percentile. IQR: Inter-Quartil Range, 25-75%. SE: Standard Error. BIBLIOGRAPHY [1]. You et al, EBioMedicine, 2020 Jan;51:102574 From the blood test to reporting the result to the physician:

News Dec 17, 2021 2 min hPG80 at ASCO 2021 - Prolevcan study Plasma hPG80 circulating prograstin levels in cancer patients in Nigeria: Prolevcan study Dec 17, 2021 3 min hPG80 at ASCO 2021 - Hepatocellular carcinoma ASCO Poster Session: Prognostic value of plasma hPG80, alone or in combination with AFP, in patients with hepatocellular carcinoma Dec 17, 2021 2 min A Major Publication: The Overexpression of hPG80 (circulating progastrin) in Multiple Cancers The oncogenic and druggable hPG80 (Progastrin) is overexpressed in multiple cancers and detected in the blood of patients Dec 17, 2021 1 min Dr. Aman Chauhan MD (Markey Cancer Center) speaking about hPG80 at #GI ASCO 2020 HPG80 (Progastrin), a novel blood-based biomarker for detection of neuroendocrine neoplasms. GI ASCO / Presented Friday, January 24, 2020

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hPG80 is a biomarker for monitoring of treatment response and recurrence in Hepatocellular Carcinoma patients Contract signed with: CHU Montpellier, France Principal investigator: Eric Assenat 1 Scientific context : Hepatocellular carcinoma (HCC) is the most common primary liver cancer and the fourth leading cause of cancer death (1) . More than 800,000 new HCC cases are diagnosed annually, and more than 800,000 patients die each year (2) . It develops in a cirrhotic liver in about 90% of cases, appearing rarely on healthy livers or with non-cirrhotic chronic liver diseases (3) . Curative treatments such as hepatic resection, liver transplantation and percutaneous ablation are offered in only 30-40% of patients (4) . Therefore, the majority of patients receive palliative care to increase survival. Treatments include transarterial chemoembolization (TACE), transarterial radioembolization (TARE), systemic therapies with chemo or molecular targeted therapies (sorafenib and regorafenib). Currently, serum alpha-fetoprotein (AFP) is the most widely used marker for diagnosing HCC. However, with a cut-off value of 20 ng/mL, the sensitivity of AFP is only 60%, and therefore AFP alone should not be used for screening (5) . Indeed, AFP levels are not elevated in 80% of patients with small tumors (6, 7) . On the other hand, AFP levels can be increased in patients with chronic liver disease (e.g. hepatitis) (8) . Furthermore, the use of AFP in HCC surveillance remains controversial (9) . Progastrin is an intracellular protein that is, or not, maturated into gastrin. When progastrin is maturated into gastrin, it is released from the cells. When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. If progastrin is not maturated into gastrin, it is released from the cells as such and named hPG80. This only happens in tumor cells: progastrin becomes a circulating protein, hPG80, which can be detected in the blood of cancer patients. The expression of the progastrin gene, GAST, is frequently increased, at early stages of tumor development, especially in colorectal cancer (but also in other cancers such as stomach, pancreatic, lung or ovarian cancer) (10) . Several signaling pathways have been involved in this process. In particular, activation of the Wnt/β-catenin pathway leads to overexpression of GASTand is involved in hepatic regeneration and in the transcriptional control of the metabolic compartmentalization of hepatic functions. Three types of liver tumors are associated with an aberrant activation of the Wnt/β-catenin pathway: hepatoblastoma, HCC and hepatocellular adenoma. In this context, hPG80 dosage might be used for early diagnosis of HCC. 2 hPG80, a new blood based biomarker Non Pathologic Condition Progastrin is not found in the blood of healthy people. When progastrin is maturated into gastrin, it is released from the cells. ​ When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. Objective Pathologic Condition hPG 80 is detected in the blood of cancer patients. When progastrin is not maturated into gastrin, it is released from the cells as such and named hPG 80 . ​ This only happens in tumor cells, whatever the tumor cell: progastrin becomes a circulating protein, hPG 80 , which can be detected in the blood of cancer patients. 3 Objectives: ​ Evaluate the value of hPG80 blood levels in monitoring of treatment response and recurrence in hepatocellular carcinoma patients. Examine whether hPG80 outperforms AFP to diagnose and monitor the disease. Analyze whether hPG80 levels were influenced by inflammation, assessed by CRP concentration. ​ 4 Patients: The hepatocellular carcinoma (HCC) cohort (PRO-HCC) came from the CHU (Centre Hospitalier Universitaire) Montpellier biobank (BB-0033-00031; the “Liverpool” collection; DC 2014-2328; AC 2014-2335; Montpellier, France). PRO-HCC is a cohort of 84 patients with HCC, managed with local or systemic treatments (nevaxar, tepotinib, regorafenib, nivolumab, anti-FGR or carbozantinib), including molecular targeted agents (“Liverpool” collection). 5 Results: 5. 1 hPG80 is detected at all stages Figure 2. hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort at progression vs remission. (A) All patients (n=84): Changes in median hPG80 levels from baseline (11.54 pM (IQR: 3.25 pM-28.28 pM)), to progression (15.71 pM (IQR:6.33-37.26 pM)), or remission (1.99 pM (IQR: 0.00-8.30 pM)). (B) Patients with normal alfa-fetoprotein (AFP) value (n=32): Changes in median hPG80 levels from baseline (14.16 pM (IQR: 7.56 pM-42.34 pM)), to progression (18.33 pM (IQR: 11.71 pM-53.70 pM)), or remission (1.47 pM (IQR: 0.21 pM-4.44 pM)). Figure 1. ​ hPG80 levels at different disease stages (focal, n=23; locally advanced, n=42; metastatic, n=19) and at disease remission after treatment (n=32). In order to simplify the reading of the graph, only statistically significant differences were shown on the graph. All the other comparisons were tested and none of them were significant. The study cohort comprises 84 patients with HCC at different disease stages: focal (n=23); locally advanced (n=42), metastatic (n=19) and at disease remission after treatment (n=32). ​ As shown in Figure 1 and 2A, hPG80 was detected in the blood of HCC patients whatever the stages. Patients in remission after disease management had lower hPG80 levels compared to those with active cancers. 5. 2 Comparison between hPG80 and AFP Figure 3. hPG80 and AFP levels in all HCC patients. 5. 3 Diagnostic performance of hPG80 in HCC patients Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic discriminative accuracy of hPG80 levels in HCC patients compared to healthy blood donors control group. As shown on Figure 3, hPG80 levels displayed high predictive significance, with an area under the curve (AUC) value of 0.85 (95% CI: 0.79-0.91; p< 0.0001) when compared to healthy blood donors. Figure 4. Diagnostic discriminative accuracy of hPG80 in patients with HCC compared to 137 healthy blood donors (age 18-25 years old) using Receiver Operating Characteristics (ROC) curve analysis. 5. 4 hPG80 and AFP kinetics in HCC patients receiving cancer treatment Individual concentration versus time curves of hPG80 evolved consistently with disease activity and AFP kinetics in most patients. This is illustrated by seven typical patients profiles (Figure 5, 6 and 7). Figure 5. ​ hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort. Longitudinal kinetics of alfa-fetoprotein (AFP) and hPG80 in 4 typical HCC patients during treatments. Figure 6. hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort. ​ Illustrative hPG80 longitudinal changes around and during disease management (baseline; remission; progression), with associated imaging obtained at the same times (multifocal liver involvement at baseline; remission after treatment with nivolumab; new liver lesions on ultrasound at progression) in a typical patient. Figure 7. ​ hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort with imaging. Longitudinal kinetics of alfa-fetoprotein (AFP) and hPG80 during treatments in 2 typical HCC patients, with consistent imaging findings. The AFP of the patient in panel A was not informative due to low concentration below the upper limit-of-normal 20 ng/ml cut-off. 5. 5 hPG80 and AFP kinetics in HCC patients receiving cancer treatment 1 Scientific context Hepatocellular carcinoma (HCC) is the most common primary liver cancer and the fourth leading cause of cancer death (1). More than 800,000 new HCC cases are diagnosed annually, and more than 800,000 patients die each year (2). It develops in a cirrhotic liver in about 90% of cases, appearing rarely on healthy livers or with non-cirrhotic chronic liver diseases (3). Curative treatments such as hepatic resection, liver transplantation and percutaneous ablation are offered in only 30-40% of patients (4). Therefore, the majority of patients receive palliative care to increase survival. Treatments include transarterial chemoembolization (TACE), transarterial radioembolization (TARE), systemic therapies with chemo or molecular targeted therapies (sorafenib and regorafenib). Currently, serum alpha-fetoprotein (AFP) is the most widely used marker for diagnosing HCC. However, with a cut-off value of 20 ng/mL, the sensitivity of AFP is only 60%, and therefore AFP alone should not be used for screening (5). Indeed, AFP levels are not elevated in 80% of patients with small tumors (6, 7). On the other hand, AFP levels can be increased in patients with chronic liver disease (e.g. hepatitis) (8). Furthermore, the use of AFP in HCC surveillance remains controversial (9). Progastrin is an intracellular protein that is, or not, maturated into gastrin. When progastrin is maturated into gastrin, it is released from the cells. When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. If progastrin is not maturated into gastrin, it is released from the cells as such and named hPG80. This only happens in tumor cells: progastrin becomes a circulating protein, hPG80, which can be detected in the blood of cancer patients. The expression of the progastrin gene, GAST, is frequently increased, at early stages of tumor development, especially in colorectal cancer (but also in other cancers such as stomach, pancreatic, lung or ovarian cancer) (10). Several signaling pathways have been involved in this process. In particular, activation of the Wnt/β-catenin pathway leads to overexpression of GAST and is involved in hepatic regeneration and in the transcriptional control of the metabolic compartmentalization of hepatic functions. Three types of liver tumors are associated with an aberrant activation of the Wnt/β-catenin pathway: hepatoblastoma, HCC and hepatocellular adenoma. In this context, hPG80 dosage might be used for early diagnosis of HCC. 2 hPG80, a new blood based biomarker Non Pathologic Condition hPG80 is not found in the blood of healthy people. When progastrin is maturated into gastrin, it is released from the cells. ​ When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. Objective Pathologic Condition When progastrin is not maturated into gastrin, it is released from the cells as such and named hPG80. ​ This only happens in tumor cells, whatever the tumor cell: progastrin becomes a circulating protein, hPG80, which can be detected in the blood of cancer patients. hPG80 is detected in the blood of cancer patients. 3 Objectives ​ Evaluate the value of hPG80 blood levels in monitoring of treatment response and recurrence in hepatocellular carcinoma patients. Examine whether hPG80 outperforms AFP to diagnose and monitor the disease. Analyze whether hPG80 levels were influenced by inflammation, assessed by CRP concentration. ​ 4 Patients The hepatocellular carcinoma (HCC) cohort (PRO-HCC) came from the CHU (Centre Hospitalier Universitaire) Montpellier biobank (BB-0033-00031; the “Liverpool” collection; DC 2014-2328; AC 2014-2335; Montpellier, France). PRO-HCC is a cohort of 84 patients with HCC, managed with local or systemic treatments (nevaxar, tepotinib, regorafenib, nivolumab, anti-FGR or carbozantinib), including molecular targeted agents (“Liverpool” collection). 5 Result 5.1 hPG80 is detected at all stages Figure 2. hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort at progression vs remission. (A) All patients (n=84): Changes in median hPG80 levels from baseline (11.54 pM (IQR: 3.25 pM-28.28 pM)), to progression (15.71 pM (IQR:6.33-37.26 pM)), or remission (1.99 pM (IQR: 0.00-8.30 pM)). (B) Patients with normal alfa-fetoprotein (AFP) value (n=32): Changes in median hPG80 levels from baseline (14.16 pM (IQR: 7.56 pM-42.34 pM)), to progression (18.33 pM (IQR: 11.71 pM-53.70 pM)), or remission (1.47 pM (IQR: 0.21 pM-4.44 pM)). Figure 1. ​ hPG80 levels at different disease stages (focal, n=23; locally advanced, n=42; metastatic, n=19) and at disease remission after treatment (n=32). In order to simplify the reading of the graph, only statistically significant differences were shown on the graph. All the other comparisons were tested and none of them were significant. The study cohort comprises 84 patients with HCC at different disease stages: focal (n=23); locally advanced (n=42), metastatic (n=19) and at disease remission after treatment (n=32). ​ As shown in Figure 1 and 2A, hPG80 was detected in the blood of HCC patients whatever the stages. Patients in remission after disease management had lower hPG80 levels compared to those with active cancers. 5.2 Comparison between hPG80 and AFP Figure 3. hPG80 and AFP levels in all HCC patients. 5.3 Diagnostic performance of hPG80 in HCC patients Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic discriminative accuracy of hPG80 levels in HCC patients compared to healthy blood donors control group. As shown on Figure 3, hPG80 levels displayed high predictive significance, with an area under the curve (AUC) value of 0.85 (95% CI: 0.79-0.91; p< 0.0001) when compared to healthy blood donors. Figure 4. Diagnostic discriminative accuracy of hPG80 in patients with HCC compared to 137 healthy blood donors (age 18-25 years old) using Receiver Operating Characteristics (ROC) curve analysis. 5.4 hPG80 and AFP kinetics in HCC patients receiving cancer treatment Individual concentration versus time curves of hPG80 evolved consistently with disease activity and AFP kinetics in most patients. This is illustrated by seven typical patients profiles (Figure 5, 6 and 7). Figure 5. ​ hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort. Longitudinal kinetics of alfa-fetoprotein (AFP) and hPG80 in 4 typical HCC patients during treatments. Figure 6. hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort. ​ Illustrative hPG80 longitudinal changes around and during disease management (baseline; remission; progression), with associated imaging obtained at the same times (multifocal liver involvement at baseline; remission after treatment with nivolumab; new liver lesions on ultrasound at progression) in a typical patient. Figure 7. ​ hPG80 kinetics in patients receiving cancer treatments in PRO-HCC cohort with imaging. Longitudinal kinetics of alfa-fetoprotein (AFP) and hPG80 during treatments in 2 typical HCC patients, with consistent imaging findings. The AFP of the patient in panel A was not informative due to low concentration below the upper limit-of-normal 20 ng/ml cut-off. 5.5 hPG80 and AFP kinetics in HCC patients receiving cancer treatment Figure 8. Impact of CRP on hPG80 levels in patients with cancer. Baseline hPG80 concentrations versus C reactive protein levels (CRP) in the PRO-HCC cohort. As shown on Figure 8, we found no link between hPG80 and inflammation status, assessed by CRP concentration, suggesting that, if any, impact of inflammation is probably limited. 6 Conclusion hPG80 is detected in the blood of HCC patients whatever the stage and remission is associated to lower levels of hPG80. ​ Upon treatment, hG80 follows disease evolution and witnesses treatment efficacy and recurrence. ​ Therefore, these data support the potential use of hPG80 as a biomarker for HCC patient follow-up. ​ In addition, we showed that hPG80 is a better biomarker than AFP to detect HCC. 7 Bibliography 1. Yang JD, Hainaut P, Gores GJ, Amadou A, Plymoth A, Roberts LR.2019. A global view of hepatocellular carcinoma: trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol 16:589-604. ​ 2. Global Burden of Disease Liver Cancer C, Akinyemiju T, Abera S, Ahmed M, Alam N, Alemayohu MA, Allen C, Al-Raddadi R, Alvis-Guzman N, Amoako Y, Artaman A, Ayele TA, Barac A, Bensenor I, Berhane A, Bhutta Z, Castillo-Rivas J, Chitheer A, Choi JY, Cowie B, Dandona L, Dandona R, Dey S, Dicker D, Phuc H, Ekwueme DU, Zaki MS, Fischer F, Furst T, Hancock J, Hay SI, Hotez P, Jee SH, Kasaeian A, Khader Y, Khang YH, Kumar A, Kutz M, Larson H, Lopez A, Lunevicius R, Malekzadeh R, McAlinden C, Meier T, Mendoza W, Mokdad A, Moradi-Lakeh M, Nagel G, Nguyen Q, Nguyen G, et al.2017. The Burden of Primary Liver Cancer and Underlying Etiologies From 1990 to 2015 at the Global, Regional, and National Level: Results From the Global Burden of Disease Study 2015. 3:1683-1691. ​ 3. Renedo F DlRJ, Calleja JL.2008. Carcinoma hepatocelular. Medicine 10:770–6. ​ 4. Bruix J, Llovet JM.2002. Prognostic prediction and treatment strategy in hepatocellular carcinoma. Hepatology 35:519-24. ​ 5. Trevisani F, D'Intino PE, Morselli-Labate AM, Mazzella G, Accogli E, Caraceni P, Domenicali M, De Notariis S, Roda E, Bernardi M.2001. Serum alpha-fetoprotein for diagnosis of hepatocellular carcinoma in patients with chronic liver disease: influence of HBsAg and anti-HCV status. J Hepatol 34:570-5. ​ 6. Zhang XF, Qi X, Meng B, Liu C, Yu L, Wang B, Lv Y.2010. Prognosis evaluation in alpha-fetoprotein negative hepatocellular carcinoma after hepatectomy: comparison of five staging systems. Eur J Surg Oncol 36:718-24. ​ 7. Agopian VG, Harlander-Locke MP, Markovic D, Zarrinpar A, Kaldas FM, Cheng EY, Yersiz H, Farmer DG, Hiatt JR, Busuttil RW.2017. Evaluation of Patients With Hepatocellular Carcinomas That Do Not Produce alpha-Fetoprotein. JAMA Surg 152:55-64. ​ 8. Toyoda H, Kumada T, Kiriyama S, Sone Y, Tanikawa M, Hisanaga Y, Hayashi K, Honda T, Kitabatake S, Kuzuya T, Nonogaki K, Kasugai T, Shimizu J.2004. Changes in the characteristics and survival rate of hepatocellular carcinoma from 1976 to 2000: analysis of 1365 patients in a single institution in Japan. Cancer 100:2415-21. ​ 9. Song PP, Xia JF, Inagaki Y, Hasegawa K, Sakamoto Y, Kokudo N, Tang W.2016. Controversies regarding and perspectives on clinical utility of biomarkers in hepatocellular carcinoma. World J Gastroenterol 22:262-74. ​ 10. You B, Mercier F, Assenat E, et al : The oncogenic and druggable hPG80 (Progastrin) is overexpressed in multiple cancers and detected in the blood of patients. EBioMedicine 51:102574, 2020

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Press Corner PRESS ENQUIRIES ➜ Contact us via e-mail Subscription Welcome to our press mailing list OUR HISTORY ➜ From the lab to the patient PHOTOS & LOGOS PHOTOS & LOGOS PRESS RELEASES VIDEOS VIDEOS PRESS RELEASES Dec 17, 2021 2 min hPG80 at ASCO 2021 - Prolevcan study Plasma hPG80 circulating prograstin levels in cancer patients in Nigeria: Prolevcan study Dec 17, 2021 3 min hPG80 at ASCO 2021 - Hepatocellular carcinoma ASCO Poster Session: Prognostic value of plasma hPG80, alone or in combination with AFP, in patients with hepatocellular carcinoma PHOTOS PRESS RELEASES PRESS RELEASES VIDEOS VIDEOS PHOTOS & LOGOS Icone Logo format: png Biodena Care logo VIDEOS PHOTOS & LOGOS PHOTOS & LOGOS VIDEOS PRESS RELEASES PRESS RELEASES Dominique Joubert Presentation Video length : 2:30' French (English subtitle) Royalty-free video on demand ​ From the lab to the patient Performing a scientific demonstration and bringing it to the patient is the dream of every scientist that Dominique Joubert is living. The scientific demonstration in question here is a major precision: to be able to detect cancer and to reverse the tumour process, in other words, to "bring back to normal cells that were cancerous". Our Montpellier laboratory Video length : 1:01' Video without audio royalty-free video on demand ​ ​ Lyon CHU laboratory Video length : 3:06' Video without audio royalty-free video on demand ​ ​ It's time to control cancer Contact us Send Your message has been sent. Contact Press

Quantification of hPG80 in the blood of patients with adenomatous and hyperplastic polyps and early stage colorectal cancer Contract signed with: Montpellier Cancer Institute, France ​ Principal investigator: Marc Ychou Liquid biopsy will become the new standard for diagnosing tumors and monitoring treatments (Bardelli et al, Cancer Cell, 2017). Plasma hPG80 detection assay falls within this framework. ECS-Progastrin has developed a sandwich ELISA assay, which is reliable, simple, robust, inexpensive and used in almost all laboratories and hospitals around the world. ​ Using our ELISA assay, we have already demonstrated the presence of hPG80 on a first cohort of colorectal cancer patients, mainly patients in late stages (more than 150 patients in stage 3 and 4) and some early stages (20 patients in stage 1 and 2). Whatever the stage, hPG80 is detected with very good sensitivity. Similarly, we showed the presence of hPG80 in the blood of 58 patients with adenomatous polyps, with very good sensitivity. With regard to hyperplastic polyps, we were only able to analyze the presence of hPG80 in a limited number of cases (n = 10). hPG80 was not detected in any of these patients. ​ This project aims to increase the number of patients with stage I and II colorectal cancer or with hyperplastic or adenomatous polyps, for whom we will measure plasma hPG80 levels. This will clarify the value of hPG80 as a biomarker for patients with early stages colorectal cancer and for patients with polyps. Non Pathologic Condition hPG80 is not found in the blood of healthy people. When progastrin is maturated into gastrin, it is released from the cells. ​ When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. Pathologic Condition hPG80 is detected in the blood of cancer patients. When progastrin is not maturated into gastrin, it is released from the cells as such and named hPG80. ​ This only happens in tumor cells, whatever the tumor cell: progastrin becomes a circulating protein, hPG80, which can be detected in the blood of cancer patients. Show the presence or absence of hPG80 in the blood of patients with adenomatous polyps or with early colorectal cancer and establish the discriminative accuracy of hPG80 as a biomarker of these pathologies. ​ Correlate hPG80 levels with cancer stage. Cancer patients samples were obtained from SIRIC (Montpellier, FRANCE). Analyzes were performed within Eurobiodev facility (Montpellier, FRANCE). This retrospective study was approved by Montpellier cancer institute scientific council (ICM-CORT). Approval date and number: 13/11/2017 (ICM-CORT-2017-23) 96 patients were included in the study. Healthy blood donors samples were obtained from the French blood agency (EFS, Montpellier, FRANCE). Analyzes were performed within Eurobiodev facility (Montpellier, FRANCE). Approval date and number: 17/07/2017 (21PLER2017-0019) 80 non-fasting healthy donors were included in the study. The presence of hPG80 in early colorectal cancer stages was confirmed. ​ Median hPG80 levels in CRC early stages patients were significantly higher than those in healthy blood donors (p< 0.0001) (Figure 1). Median hPG80 concentration was 0.23 pM (IQR 0.00-0.80 pM) in control population, against 2.315 pM (IQR 0.64-3.69 pM), 2.26 pM (IQR 0.70-9.60 pM), and 5.46 pM (IQR 1.67-9.945 pM) in patients with preneoplasic lesions, CRC stage 1 or CRC stage 2, respectively (Figure 1). There is a significant increase in hPG80 blood levels from adenomatous polyps to stage 2 colorectal cancer. Figure 1. Comparison of hPG80 concentrations between healthy blood donors (n=80) and cancers patients, preneoplasic lesions (n=24), stage 1 CRC (n=23), stage 2 CRC (n=49). Representation in box and Whiskers (2.5-97.5 percentile, dots are outliers). Two-tailed Mann-Whitney test; ***, p< 0.001; *, p< 0.05. The diagnostic discriminative accuracy of hPG80 levels was estimated using Receiver Operating Characteristics (ROC) curves in patients with preneoplasic lesions, patients with stage 1 CRC or patients with stage 2 CRC, compared to healthy blood donors control cohort. ​ The diagnostic discriminative accuracy estimated by the ROC AUC were 0.80 (95% CI: 0.69 to 0.91) for patients with preneoplasic lesions; 0.825 (95% CI: 0.72 to 0.93) for patients with stage 1 CRC; and 0.89 (95% CI: 0.82 to 0.95) for patients with stage 2 CRC, compared to the blood donor control group (Figures 2, 3 and 4). 1 Scientific context 2 hPG80, a new blood based biomarker 3 Objectives 4 Patients 5 Results 6 Conclusion hPG80 is detected with a very good accuracy in the blood of patients with an adenomatous polyp or with a stage 1 or 2 colorectal cancer.

Nouveau titre Contract signed with: Montpellier Cancer Institute, France ​ Principal investigator: Marc Ychou Liquid biopsy will become the new standard for diagnosing tumors and monitoring treatments (Bardelli et al, Cancer Cell, 2017). Plasma hPG80 detection assay falls within this framework. ECS-Progastrin has developed a sandwich ELISA assay, which is reliable, simple, robust, inexpensive and used in almost all laboratories and hospitals around the world. ​ Using our ELISA assay, we have already demonstrated the presence of hPG80 on a first cohort of colorectal cancer patients, mainly patients in late stages (more than 150 patients in stage 3 and 4) and some early stages (20 patients in stage 1 and 2). Whatever the stage, hPG80 is detected with very good sensitivity. Similarly, we showed the presence of hPG80 in the blood of 58 patients with adenomatous polyps, with very good sensitivity. With regard to hyperplastic polyps, we were only able to analyze the presence of hPG80 in a limited number of cases (n = 10). hPG80 was not detected in any of these patients. ​ This project aims to increase the number of patients with stage I and II colorectal cancer or with hyperplastic or adenomatous polyps, for whom we will measure plasma hPG80 levels. This will clarify the value of hPG80 as a biomarker for patients with early stages colorectal cancer and for patients with polyps. Non Pathologic Condition hPG80 is not found in the blood of healthy people. When progastrin is maturated into gastrin, it is released from the cells. ​ When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. Pathologic Condition hPG80 is detected in the blood of cancer patients. When progastrin is not maturated into gastrin, it is released from the cells as such and named hPG80. ​ This only happens in tumor cells, whatever the tumor cell: progastrin becomes a circulating protein, hPG80, which can be detected in the blood of cancer patients. Show the presence or absence of hPG80 in the blood of patients with adenomatous polyps or with early colorectal cancer and establish the discriminative accuracy of hPG80 as a biomarker of these pathologies. ​ Correlate hPG80 levels with cancer stage. Cancer patients samples were obtained from SIRIC (Montpellier, FRANCE). Analyzes were performed within Eurobiodev facility (Montpellier, FRANCE). This retrospective study was approved by Montpellier cancer institute scientific council (ICM-CORT). Approval date and number: 13/11/2017 (ICM-CORT-2017-23) 96 patients were included in the study. Healthy blood donors samples were obtained from the French blood agency (EFS, Montpellier, FRANCE). Analyzes were performed within Eurobiodev facility (Montpellier, FRANCE). Approval date and number: 17/07/2017 (21PLER2017-0019) 80 non-fasting healthy donors were included in the study. The presence of hPG80 in early colorectal cancer stages was confirmed. ​ Median hPG80 levels in CRC early stages patients were significantly higher than those in healthy blood donors (p< 0.0001) (Figure 1). Median hPG80 concentration was 0.23 pM (IQR 0.00-0.80 pM) in control population, against 2.315 pM (IQR 0.64-3.69 pM), 2.26 pM (IQR 0.70-9.60 pM), and 5.46 pM (IQR 1.67-9.945 pM) in patients with preneoplasic lesions, CRC stage 1 or CRC stage 2, respectively (Figure 1). There is a significant increase in hPG80 blood levels from adenomatous polyps to stage 2 colorectal cancer. Figure 1. Comparison of hPG80 concentrations between healthy blood donors (n=80) and cancers patients, preneoplasic lesions (n=24), stage 1 CRC (n=23), stage 2 CRC (n=49). Representation in box and Whiskers (2.5-97.5 percentile, dots are outliers). Two-tailed Mann-Whitney test; ***, p< 0.001; *, p< 0.05. The diagnostic discriminative accuracy of hPG80 levels was estimated using Receiver Operating Characteristics (ROC) curves in patients with preneoplasic lesions, patients with stage 1 CRC or patients with stage 2 CRC, compared to healthy blood donors control cohort. ​ The diagnostic discriminative accuracy estimated by the ROC AUC were 0.80 (95% CI: 0.69 to 0.91) for patients with preneoplasic lesions; 0.825 (95% CI: 0.72 to 0.93) for patients with stage 1 CRC; and 0.89 (95% CI: 0.82 to 0.95) for patients with stage 2 CRC, compared to the blood donor control group (Figures 2, 3 and 4). 1 Scientific context 2 hPG80, a new blood based biomarker 3 Objectives 4 Patients 5 Results 6 Conclusion hPG80 is detected with a very good accuracy in the blood of patients with an adenomatous polyp or with a stage 1 or 2 colorectal cancer.

Last Update : Feb 02, 2020 News: Events Dec 17, 2021 2 min hPG80 at ASCO 2021 - Prolevcan study Plasma hPG80 circulating prograstin levels in cancer patients in Nigeria: Prolevcan study Dec 17, 2021 3 min hPG80 at ASCO 2021 - Hepatocellular carcinoma ASCO Poster Session: Prognostic value of plasma hPG80, alone or in combination with AFP, in patients with hepatocellular carcinoma Dec 17, 2021 2 min A Major Publication: The Overexpression of hPG80 (circulating progastrin) in Multiple Cancers The oncogenic and druggable hPG80 (Progastrin) is overexpressed in multiple cancers and detected in the blood of patients Dec 17, 2021 1 min Dr. Aman Chauhan MD (Markey Cancer Center) speaking about hPG80 at #GI ASCO 2020 HPG80 (Progastrin), a novel blood-based biomarker for detection of neuroendocrine neoplasms. GI ASCO / Presented Friday, January 24, 2020

Contract signed with: Les Hospices Civils de Lyon ​ Principal investigator: Benoît You hPG 80 is a biomarker for patients treated for gastro-intestinal cancers. A retrospective study on the BIG-RENAPE cohort. Last updated: August 3, 2020 1 Scientific context There is a need for early predictor of clinical benefits able to compress the time line required to evaluate new treatments ( 1) . Moreover, peritoneal carcinomatosis frequently developed by gastrointestinal cancer patients is poorly assessable using conventional imaging ( 2) . In addition, the traditional morphological RECIST criteria were found not adequate for assessing treatment efficacy with anti-angiogenic drugs ( 3) . Strategies that may help characterize tumor size changes or treatment efficacy during treatment are to be developed. If some serum markers (liquid biopsy) were shown to be surrogate markers of treatment efficacy, they could be used to identify the patients who benefit the most from the treatment or who may be candidates for iterative surgery ( 4) . 2 hPG80, a new blood based biomarker Non Pathologic Condition hPG 80 is not found in the blood of healthy people. When progastrin is maturated into gastrin, it is released from the cells. ​ When gastrin is produced by the G cells of the stomach antrum, it plays its role to control acidic secretions during digestion. Pathologic Condition hPG 80 is detected in the blood of cancer patients. When progastrin is not maturated into gastrin, it is released from the cells as such and named hPG 80 . ​ This only happens in tumor cells, whatever the tumor cell: progastrin becomes a circulating protein, hPG 80 , which can be detected in the blood of cancer patients. 3 Objective Assessment of hPG80 usability in follow up with surgery. 4 Patients Patients recorded in prospective BIG-RENAPE project database, NCT02823860 (Lyon, FRANCE). Biological data were extracted from the data base. Analysis was performed in HCL Lyon Sud facility (Pierre-Bénite, FRANCE). 194 patients with peritoneal involvements from gastrointestinal cancers, treated with peri-operative chemotherapy regimens and cytoreductive surgery. ​ Blood samples were collected at inclusion (n=194) and between 8 and 24 hours after cytoreductive surgery, performed with or without hyperthermic intraperitoneal chemotherapy (HIPEC) (n=85). 5 Results In this cohort, median hPG80 significantly decreased from inclusion to the post-operative period from 3.08 to 1.57 pM[AP1] , p<0.0001 with two-tailed unpaired Mann-Whitney test. ​ The individual curves showed that 74.2% of patients had hPG80 titer declines >25%. The remaining patients experienced stable concentrations (16.1% of patients), or hPG80 level increases >25% (9.7% of patients). 6 Conclusion hPG80 is a good biomarker for peritoneal carcinomatosis patients to assess surgery efficacy. Levels are significantly decreased after surgery and individual follow-up shows that minimal residual disease might be questioned via the presence of hPG80 in the blood. ​ These results have been published in You et al, EBioMedicine, 2020 . 7 Bibliography 1 . Kola I, Landis J: Can the pharmaceutical industry reduce attrition rates? Nat Rev Drug Discov 3:711-5, 2004. 2 . Laghi A, Bellini D, Rengo M, et al: Diagnostic performance of computed tomography and magnetic resonance imaging for detecting peritoneal metastases: systematic review and meta-analysis. Radiol Med 122:1-15, 2017. ​ 3 . Nishino M, Jagannathan JP, Krajewski KM, et al: Personalized tumor response assessment in the era of molecular medicine: cancer-specific and therapy-specific response criteria to complement pitfalls of RECIST. AJR Am J Roentgenol 198:737-45. ​ 4 . McMullen JRW, Selleck M, Wall NR, et al: Peritoneal carcinomatosis: limits of diagnosis and the case for liquid biopsy. Oncotarget 8:43481-43490, 2017. ​ 5 . You B, Mercier F, Assenat E, et al : The oncogenic and druggable hPG80 (Progastrin) is overexpressed in multiple cancers and detected in the blood of patients. EBioMedicine 51:102574, 2020.