Specific Aims: Impact of Locoregional Therapy of Heterogenicity HCC and Tumor Biology

The liver is the second largest (after the skin) and critical organ in the body, and about 70 to 80 percent of the liver is made up of parenchymal hepatocytes, while nonparenchymal cells make up about thirty to forty percent of the liver. The liver sinusoids are comprised of sinusoidal endothelial cells (EC), and macrophages, known also as Kupffer cells, are in close contact with EC inside the sinusoids, while hepatic stellate cells exist outside the sinusoids. Additionally, the liver is a unique organ due to its dual blood supply from the portal vein (approximately 75%) and the hepatic artery (approximately 25%). I am exploring the role of cancer stem cell markers and hypoxia markers that are resistant to the tumor and CSC type HCC after TACE by addressing the followings aims:

• Aim I: To test that treated HCC express higher levels of CSCs Markers its target genes than TARE-treated HCC tissues samples from 1) TARE-treated, 2) TACE-treated 3) no treated patients (K19, CD24, EpCAM, CD133) validated by IHC

• Aim II: will test the hypothesis that TACE-treated HCC express higher levels of hypoxia markers and its target gene than TARE-treated HCC tissues samples from TARE-treated, 2) TACE-treated 3) no treated patients (CAIX, ALDOA, RITI, LOX) validated by IHC, anhydrase-IX), ALDOA (Fructose-bisphosphate aldolase), LOX (Lysyl oxidase), FAP (Fibroblast activation protein), RITI (Ras-like-without-CAAX-1)

The liver has four major functions, including synthetic function, detoxification, bile production, and immunological functions. Synthetic function includes anabolism in carbohydrate metabolism, such as gluconeogenesis glycogen synthesis; protein synthesis, such as production of albumin; and lipid metabolism, such as production of lipoprotein and cholesterol synthesis. Detoxification involves catabolism and degradation of multiple endogenous and exogeneous products, such as bilirubin, ammonia, and drugs. Bile production/secretion is directly involved in digestion and absorption of lipids, and immunological functions include the activation of KC, which are the largest pool of resident macrophages in the body.

The liver is the target of many injuries due to the hard work it does to detoxify and keep the body safe. Some these injuries are temporary, but others are more chronic and persistent, such as chronic viral hepatitis, which is one of the common chronic inflammatory conditions of the liver. Fibrosis, another chronic and persistent disease that causes liver injuries, can arise due to alcoholism, excessive fat in the liver, and metabolic defects that results in chronic damage of the hepatocytes (Author, year, page #). Cirrhosis, the combination of advanced fibrosis and abnormal hepatic nodular regeneration, is histologically of combination of advanced liver fibrosis and hepatic regeneration/hepatic regenerative nodules. It stops the liver from working normally in advanced stages, which causes major liver function impairment, and it also causes an increased risk of hepatocellular carcinoma (HCC) (Author, year, page #). Cirrhosis alone or combined with HCC, can ultimately cause organ failure and death (Author, year, page #).

My experimental designs include Paraffin-embedded specimens obtained from a cohort of HCC patients after surgery. I will be using HCC tissue samples from 3 main groups: TACE treated, TARE treated and no treatment patient (Tumor, Non-tumor). The majority of the patients are HCV infected with more frequent in male than female. I am exploring my aims by performing standard IHC. Staining slides were imaged using a Leica DM light microscope and a SPOT Insight firewire 4 mega sample camera. I normally take between 5-7 photos and choose the best five, with lower background signaling noise.

Currently, I have finished all my staining for the 18 patients both tumor and non-tumor sections for my stemness markers, and I am working on an analysis of cancer stem cell markers previously identified in metastatic prostate cancer patients. I need to work on taking the image and analysis for the following markers (CD133,K19), and I am planning to finish that by end of October. November -February 2020 I am planning to work on my second aim which is the hypoxia markers, taking photos of the slides and do the analysis.

References

Demetris, A. J., et. al. (2016). Functional immune anatomy of the liver-as an allograft. Am JTransplant, 16(6), 1653–1680.

Fujiwara, N., et al. (2018). Risk factors and prevention of hepatocellular carcinoma in the era of precisionmedicine. J Hepatol, 68(3), 526-549.

Michalopoulos, G.K. (2017). Hepatostat: Liver regeneration and normal liver tissue maintenance. Hepatology, 65(4), 1384-1392.

Sukowati, C. (2019). Heterogeneity of hepatic cancer stem cells. Adv Exp Med Biol, 1139,59-81.

Yun, Z and Lin, Q. (2014). Hypoxia and regulation of cancer cell stemness. Adv Exp Med Biol,772, 41-53.

Zhu, S. and Hoshida, Y. (2018). Molecular heterogeneity in hepatocellular carcinoma. Hepat Oncol, 5(1).