Cancer is an abnormal growth of cells which has the potential to spread to other parts of the body and destroy normal body tissue. Cancer is a major threat to health representing the second largest cause of mortality in the world following cardiovascular diseases, but thanks to advancement in cancer screening and treatment, survival rates are constantly increasing for different types of cancer. The etiology of cancer is still being investigated worldwide. In general, most cancers associated with DNA mutations, where the mutation can cause a healthy cell to allow rapid cell growth, fail to stop uncontrolled cell growth, or make mistakes when repairing DNA errors.  Age, habit, family history, environmental factors, lifestyle, certain health conditions, including ulcerative colitis are common factors known to increase the risk of developing cancer. According to the WHO, the number of global deaths is expected to rise by 45% between 2008 and 2030, and by 2030 between 10 million and 11 million cancers will be diagnosed each year.

Dating as far back as the 1900’s mass spectrometry is one of the most widely used analytical methods in biosciences and medical research. Mass spectrometry-based proteomics was developed as a primary tool for the identification of medically relevant biomarkers for disease advancement and analyzing response to treatment with better results for patients. Since its advent, this analytical technique has advanced steadily, being utilized in various applications ranging from forensic toxicology to cancer diagnosis.

Several scientific papers published in 2020 including those by Uzzaman et al [1] and Sun et al [2] describe the use of mass spectrometers in cancer research. Also, Liu et al [3] used a combination of MS-intensive methods such as isobaric tags for relative and absolute quantitation with two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2D LC-MS/MS) and 1D-targeted LC-MS/MS, on serum samples from healthy people (normal control, NC), patients with benign diseases (BD), and Pancreatic Cancer (PC) patients to identify novel biomarkers of PC. They identified more than 1000 proteins, verified 142 differentially expressed proteins, and finally targeted four proteins for absolute quantitation in 100 serum samples. The novel biomarker panel of apolipoprotein E (APOE), inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3), apolipoprotein A-I (APOA1), apolipoprotein L1 (APOL1), combining with CA19-9, statistically-significantly improved the sensitivity (95%) and specificity (94.1%), outperforming CA19-9 alone, for the diagnosis of PC. Radu et al [2] reports that mass spectrometry provides a powerful and sensitive approach for the analysis of biological systems, and dedicated instruments are available for different classes of analytes, such as proteomes, metabolomes, genomes, transcriptomes, etc.

ITSI-Biosciences offers comprehensive mass spectrometry services for cancer research. Specifically, services such as protein identification, protein quantitation, biomarker discovery, global protein expression profiling and post translational modification (PTM) mapping can be applied to all areas of cancer research. ITSI has been offering such mass spec services since 2005. Hundreds of human and non-human samples have been analyzed and thousands of proteins and candidate biomarkers have been identified by mass spectrometry. As little as 1ug of total protein can be analyzed. There are several complementary technologies including Agilent Bioanalyzer, Luminex xMAP and GE 2D-DIGE platforms are available at ITSI to support mass spectrometry-based cancer research. Figure 1 is a generalized ITSI LC-MS/MS workflow which has been applied to the study of several cancers [4-9].

Figure 1: Generalized workflow for biological sample analysis by liquid chromatography – tandem mass spectrometry (LC-MS/MS) at ITSI. Post translational modification mapping (PTM) and gene ontology classification are additional services that add value to the data obtained by mass spectrometry.

ITSI Biosciences uses LC-MS/MS ion trap mass spectrometry technology.  All biological sample types can be analyzed at  ITSI-Biosciences, including 1D-gel slices, 2D gel spots, protein lysate, blood, serum, plasma, saliva, urine, whole tissue, western blot membranes, formalin-fixed paraffin embedded samples and laser micro-dissected samples. The standard mass spectrometry workflow may involve processing samples in a specialized way to isolate proteins, the proteins are reduced and alkylated. The samples then go through a cleanup step using our in-house ToPREP total protein precipitation kit.  The precipitated samples are resuspended in a digestion buffer and digested with trypsin or other enzyme as needed to achieve the goals of the analysis. The digested peptides are then separated by an inline liquid chromatography step at a nanoliter flow rate, to separate the peptides by reverse phase chromatography on a C18 column prior to an electrospray ionization step.  The ionized peptides are then analyzed using a high performance LTQ XL or Q Exactive orbitrap mass spectrometer.  The raw data files are further processed using Proteome Discoverer to identify peptides, proteins, and common peptide modifications. Additionally, Gene Ontology can be performed to define the molecular functions, cellular locations and biological process that the identified proteins may carry out.

The typical turnaround is 1-2 weeks depending on the project. For more information, contact ITSI-Biosciences via Email; itsi@itsibio.com, Phone; 814-262-7331 or Fax; 814-262-7334.

Reference

  1. Uzzaman A, Zhang X, Qiao Z, Zhan H, Sohail A, Wahid A, Shang Z, Guan X, Cao CX, Xiao H (2020). Discovery of small extracellular vesicle proteins from human serum for liver cirrhosis and liver cancer. 2020 Aug 21:S0300-9084(20)30198-X. doi: 10.1016/j.biochi.2020.08.013. Online ahead of print.PMID: 32835735
  2. Sun S, Zhang H, Wang Y, Gao J, Zhou S, Li Y, Han S, Li X, Li J (2020). Proteomic Analysis of Human Esophageal CancerUsing Tandem Mass Tag Quantifications. Biomed Res Int. 2020 Aug 7;2020:5849323. doi: 10.1155/2020/5849323. eCollection 2020.PMID: 32832552
  3. Liu X, Zheng W, Wang W, Shen H, et al (2017). A new panel of pancreatic cancer biomarkers discovered using a mass spectrometry-based pipeline. Br J Cancer. 2017 Dec 5; 117(12): 1846–1854. doi: 1038/bjc.2017.365.
  4. Radu Albulescu, Andrei Jose Petrescu, Mirela Sarbu, et al (2019). Mass Spectrometry for Cancer Biomarkers, Proteomics Technologies and Applications, Ibrokhim Y. Abdurakhmonov, IntechOpen, DOI: 10.5772/intechopen.85609. Available from: https://www.intechopen.com/books/proteomics-technologies-and-applications/mass-spectrometry-for-cancer-biomarkers
  5. Somiari RI, Sullivan, A, Russell, S, Somiari, S, Hu, H, Jordan, R, George, A, Katenhusen, R, Buchowiecka, A, Arciero, C, Brzeski, H, Hooke, J, Shriver, C. (2003). High throughput proteomic analysis of infiltrating ductal carcinoma of the breast. Journal of Proteomics. 10 (3): 1863 – 1873
  6. Somiari, RI, Somiari SB, Russell S and Shriver CD. (2005) Proteomics of breast carcinoma. J. Chromatography B, Analyt Technol Biomed Life Sci. 815: 215-225.
  7. Boyiri T, Somiari RI, Russell, S, Aliaga, C and El-Bayoumy K (2009). Proteomics of rat prostate lobes treated with 2-N-hydroxylamino-1-methyl-6-phenylimidazo [4,5-b] pyridine and 5α-dihydrotestosterone. International Journal of Oncology 35: 559-567.
  8. Bortner J; Das A; Umstead T; Freeman W; Somiari R; Aliaga C; Phelps D; El-Bayoumy K (2009). Down-Regulation of 14-3-3 Isoforms and Annexin A5 Proteins in Lung Adenocarcinoma – Induced by the Tobacco-Specific Nitrosamine NNK in the A/J Mouse Revealed by Proteomic Analysis. Journal of Proteome Research 8(8): 4050-4061
  9. Arciero C, Somiari, S, Shriver C, Brzeski, H, Jordan, R, Hu H, Ellsworth D, Somiari, RI (2003). Functional relationship and gene ontology classification of breast cancer biomarkers. International Journal of Biological Markers 18 (4): 241-272.