Novel Biomarkers of Hypoxia and Metabolism in Clear Cell Renal Cell Carcinoma
Study Details
Study Description
Brief Summary
Renal cell carcinoma (RCC) is one of the common malignant tumors in human beings and originates from the renal tubular epithelium. Clear cell renal cell carcinoma (ccRCC) is the main pathological type of RCC. Due to the lack of reliable biomarkers and clinical symptoms for early diagnosis, imaging findings such as ultrasound and CT are needed. When the patients presented typical symptoms, for example, hematuria, backache, and abdominal mass, some of them are in advanced stages of cancer. About a quarter of patients had metastasis at the first diagnosis, and the 5-year survival rate of these patients was less than 10%. Therefore, the early diagnosis of ccRCC and the prevention of tumor recurrence and metastasis are of great significance.
The preliminary studies suggested that some hypoxia and metabolism-related molecules were highly expressed in ccRCC tumors but low in normal kidney tissues. The molecules included carbonic anhydrase IX/9 (CA IX/CA9), the mitochondrial NADH dehydrogenase [ubiquinone] 1 alpha subcomplex, 4-like 2(NDUFA4L2), angiopoietin-like protein 4(ANGPTL4), hypoxia inducible lipid droplet-associated (HILPDA), and egl-9 family hypoxia-inducible factor 3( EGLN3) et al . Cell-free DNA methylomes were also highly expressed in the blood of ccRCC patients.
In order to further verify the expression status of the above novel biomarkers in ccRCC, the investigators will detect the expressions of these molecules in the tumor and adjacent tissues from 140 ccRCC patients by RT-PCR, Western blot, and immunohistochemistry.140 healthy people were selected as the control group. 30 patients with benign kidney diseases were selected as another control group. Blood and urine samples from the ccRCC group and the control group were collected. The mRNA and protein levels of the above molecules in blood or urine samples were detected by qRT-PCR and ELISA. The correlation between the expression of the above new biomarkers and clinical data, such as early diagnosis, pathological grade, recurrence and metastasis, and survival time, was statistically analyzed. The above molecular changes were dynamically detected before surgery, 1 week, and 6 months after surgery. A receiver-operating characteristic curve (ROC) was used to determine the threshold value of these biomarkers for the diagnosis of renal clear cell carcinoma.
The study is to explore the specific tumor biomarker spectrum for clinical diagnosis, evaluation of recurrence, metastasis, and prognosis of ccRCC, which will be auxiliary early screening and diagnosis, reducing the harm of renal cancer to human health.
Condition or Disease | Intervention/Treatment | Phase |
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Study Design
Arms and Interventions
Arm | Intervention/Treatment |
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the ccRCC patient group The age and gender of patients are not limited. The patient was preliminarily diagnosed as renal carcinoma by imaging examination and finally diagnosed as clear cell renal cell carcinoma by pathology. |
Diagnostic Test: No intervention was required for patients or control group in this study
The patient was treated normally and no intervention was required in this study
|
the healthy control group The age and sex of the healthy control group were matched with that of ccRCC patient group. There was no tumor in the kidney or other parts of the body, and no tumor in the blood system. The healthy control group did not have any renal benign diseases, such as kidney stones, diabetic nephropathy, inflammation, and uremia. There are no inflammatory diseases in other parts of the body. The functions of the liver, kidney, and heart were normal. |
Diagnostic Test: No intervention was required for patients or control group in this study
The patient was treated normally and no intervention was required in this study
|
the benign kidney disease group The age and sex of this group were matched with that of ccRCC patient group. The patient did not have any tumor but had one of the benign kidney diseases, such as kidney stones, diabetic nephropathy, inflammation, and uremia. The functions of the liver and heart were normal. |
Diagnostic Test: No intervention was required for patients or control group in this study
The patient was treated normally and no intervention was required in this study
|
Outcome Measures
Primary Outcome Measures
- Detection of novel biomarkers in ccRCC patients before surgery [For ccRCC patients, these six novel biomarkers were detected 1 to 3 days before surgery]
The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
- Detection of novel biomarkers in ccRCC patients 1 week after surgery [For ccRCC patients, the detection of these biomarkers were completed within one week after surgery]
The expressions of novel biomarkers in tissues: In the tumor and adjacent tissues from ccRCC patients, the mRNA expressions of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 were measured by real-time fluorescence quantitative PCR method. The protein expressions of these five molecules were detected by western blot, and immunohistochemistry methods. The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
- Detection of novel biomarkers in ccRCC patients at 6 months after surgery [For ccRCC patients, these six novel biomarkers were detected at 6 months postoperatively]
The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
- Detection of novel biomarkers in the control groups as baseline [For the healthy control group and the benign kidney disease group, the levels of these six biomarkers were detected as baseline.]
The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
Eligibility Criteria
Criteria
Inclusion Criteria:
- The age and sex of the healthy control group were matched with that of ccRCC patient group. There was no tumor in the kidney or other parts of the body, and no tumor in the blood system. The healthy control group did not have any renal benign diseases, such as kidney stones, diabetic nephropathy, inflammation, and uremia. There are no inflammatory diseases in other parts of the body; the functions of the liver, kidney, and heart were normal.
Exclusion Criteria:
- The volunteer has tumors in the kidney or other parts of the body, or blood system tumors; The volunteer has benign kidney diseases, such as kidney stones, diabetic nephropathy, nephritis and uremia, etc; The patient has inflammatory disease elsewhere. If the volunteer has any one of the above diseases, it shall be excluded.
Contacts and Locations
Locations
Site | City | State | Country | Postal Code | |
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1 | The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital | Jinan | China |
Sponsors and Collaborators
- Qianfoshan Hospital
Investigators
- Principal Investigator: Li Zhang, Dr., The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital
Study Documents (Full-Text)
None provided.More Information
Publications
- Galaup A, Cazes A, Le Jan S, Philippe J, Connault E, Le Coz E, Mekid H, Mir LM, Opolon P, Corvol P, Monnot C, Germain S. Angiopoietin-like 4 prevents metastasis through inhibition of vascular permeability and tumor cell motility and invasiveness. Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18721-6. Epub 2006 Nov 27.
- Liu L, Lan G, Peng L, Xie X, Peng F, Yu S, Wang Y, Tang X. NDUFA4L2 expression predicts poor prognosis in clear cell renal cell carcinoma patients. Ren Fail. 2016 Sep;38(8):1199-205. doi: 10.1080/0886022X.2016.1208517. Epub 2016 Jul 25.
- Lucarini L, Magnelli L, Schiavone N, Crisci A, Innocenti A, Puccetti L, Cianchi F, Peri S, Supuran CT, Papucci L, Masini E. Plasmatic carbonic anhydrase IX as a diagnostic marker for clear cell renal cell carcinoma. J Enzyme Inhib Med Chem. 2018 Dec;33(1):234-240. doi: 10.1080/14756366.2017.1411350.
- Miikkulainen P, Högel H, Seyednasrollah F, Rantanen K, Elo LL, Jaakkola PM. Hypoxia-inducible factor (HIF)-prolyl hydroxylase 3 (PHD3) maintains high HIF2A mRNA levels in clear cell renal cell carcinoma. J Biol Chem. 2019 Mar 8;294(10):3760-3771. doi: 10.1074/jbc.RA118.004902. Epub 2019 Jan 7.
- Moch H, Cubilla AL, Humphrey PA, Reuter VE, Ulbright TM. The 2016 WHO Classification of Tumours of the Urinary System and Male Genital Organs-Part A: Renal, Penile, and Testicular Tumours. Eur Urol. 2016 Jul;70(1):93-105. doi: 10.1016/j.eururo.2016.02.029. Epub 2016 Feb 28. Review.
- Nuzzo PV, Berchuck JE, Korthauer K, Spisak S, Nassar AH, Abou Alaiwi S, Chakravarthy A, Shen SY, Bakouny Z, Boccardo F, Steinharter J, Bouchard G, Curran CR, Pan W, Baca SC, Seo JH, Lee GM, Michaelson MD, Chang SL, Waikar SS, Sonpavde G, Irizarry RA, Pomerantz M, De Carvalho DD, Choueiri TK, Freedman ML. Detection of renal cell carcinoma using plasma and urine cell-free DNA methylomes. Nat Med. 2020 Jul;26(7):1041-1043. doi: 10.1038/s41591-020-0933-1. Epub 2020 Jun 22. Erratum in: Nat Med. 2020 Sep 7;:.
- Papandreou I, Cairns RA, Fontana L, Lim AL, Denko NC. HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. Cell Metab. 2006 Mar;3(3):187-97.
- Povero D, Johnson SM, Liu J. Hypoxia, hypoxia-inducible gene 2 (HIG2)/HILPDA, and intracellular lipolysis in cancer. Cancer Lett. 2020 Nov 28;493:71-79. doi: 10.1016/j.canlet.2020.06.013. Epub 2020 Aug 18. Review.
- Rini BI, Campbell SC, Escudier B. Renal cell carcinoma. Lancet. 2009 Mar 28;373(9669):1119-32. doi: 10.1016/S0140-6736(09)60229-4. Epub 2009 Mar 5. Review.
- Tello D, Balsa E, Acosta-Iborra B, Fuertes-Yebra E, Elorza A, Ordóñez Á, Corral-Escariz M, Soro I, López-Bernardo E, Perales-Clemente E, Martínez-Ruiz A, Enríquez JA, Aragonés J, Cadenas S, Landázuri MO. Induction of the mitochondrial NDUFA4L2 protein by HIF-1α decreases oxygen consumption by inhibiting Complex I activity. Cell Metab. 2011 Dec 7;14(6):768-79. doi: 10.1016/j.cmet.2011.10.008. Epub 2011 Nov 17.
- CT2021-11-23