Method for providing information for predicting therapeutic responsiveness or prognosis to transarterial chemoembolization

Abstract

The present invention relates to a method for providing information for predicting therapeutic responsiveness or prognosis to transarterial chemoembolization. The gut microbiota of the present invention serves as a biomarker for predicting therapeutic responsiveness or prognosis to transarterial chemoembolization in patients with liver cancer. The gut microbiota of the present invention can be utilized as a potential predictive factor for clinical outcomes of transarterial chemoembolization, can also be used as an adjunct therapeutic target for treatment of liver cancer patients, and enables prediction of survival rates of liver cancer patients who have undergone transarterial chemoembolization. Accordingly, the biomarker of the present invention is expected to be usefully applied as a marker for predicting therapeutic responsiveness or prognosis to transarterial chemoembolization in patients with liver cancer.

Description

Method for providing information for predicting treatment responsiveness or prognosis for transarterial chemoembolization

[0001] The present invention relates to a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization.

[0002] The present invention claims priority based on Korean Patent Application No. 10-2024-0138265 filed on October 11, 2024, and all contents disclosed in the specification and drawings of said applications are incorporated by reference.

[0003] The prognosis for liver cancer is extremely poor, and the possibility of treatment is limited by the severity of the liver disease and its spread into the liver. Among these, hepatocellular carcinoma (HCC) is the most common type of liver cancer in adults and accounts for the third leading cause of cancer-related death (Stefaniuk P, et al., 2010, World J Gastroenterol 16: 418-424). HCC is a disease in which symptoms only appear after it has progressed significantly; consequently, the timing for appropriate treatment is frequently missed, and even when treatment is administered, the prognosis remains extremely poor. In particular, cases where surgical resection is impossible are severe enough to result in death within one year, and significant improvements in treatment are expected if diagnostic methods are improved. Surgical resection or liver transplantation is the only treatment option for tumors and other severe liver diseases. However, these treatments are only applicable to patients with non-cirrhotic liver disease, which accounts for less than 5% of cases in Western countries. In this situation, the two most promising treatment approaches are intra-arterial or intra-tumoral metabolic radiotherapy and arterial chemoembolization with Lipiodol.

[0004] Metabolic radiation therapy is widely used for the curative or transient treatment of many cancers, particularly liver cancer. Metabolic radiation therapy for liver tumors is primarily administered via the hepatic artery. A healthy liver receives blood and oxygen simultaneously through the portal vein and hepatic artery. However, tumors, which are richly vascularized, are essentially supplied via the hepatic artery, while up to 80% of the supply to healthy tissues is delivered via the portal vein. Transarterial chemoembolization (TACE) was developed to improve the performance of metabolic radiation therapy. This technique blocks blood flow to the tumor by injecting a mixture containing chemotherapy agents combined with embolizing particles into the patient's hepatic artery. Currently, TACE is performed repeatedly, and some patients undergo unnecessary repeats despite being TACE refractory. There is currently no clear clinical definition of which patients qualify for TACE refractory. Furthermore, it exhibits various clinical manifestations, such as cases where the prognosis deteriorates due to the transformation into multiple or invasive liver cancer following repeated embolization. Therefore, a highly reliable diagnostic method is required to predict the prognosis of liver cancer chemoembolization.

[0005] Meanwhile, the microbiota (or microbiome) refers to the microbial community including bacteria, archaea, and eukaryotes present in a given habitat, and the gut microbial community is known to play an important role in human physiological processes and significantly influence human health and disease through interactions with human cells.

[0006] Metagenomics, also known as environmental genomics, can be defined as the analysis of metagenomic data obtained from samples collected from the environment. Recently, it has become possible to catalog the bacterial composition of human microbial communities using methods based on 16S ribosomal RNA (16S rRNA) sequences, and the 16S rDNA sequence, which is the gene for 16S ribosomal RNA, is analyzed using next-generation sequencing (NGS) platforms. However, there have been no reported methods for predicting the therapeutic responsiveness to transarterial chemoembolization (TACE) and the prognosis of patients with liver cancer or hepatocellular carcinoma through such metagenomic analysis.

[0007] The object of the present invention is to provide a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0008] (a) Step of separating control group and subject samples;

[0009] (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis; and

[0010] (c) If, compared to the control group in step (b) above, the content of one or more bacteria of the family Prevotellaceae and Ruminococcaceae selected from the group consisting of Prevotella and Faecalibacterium, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, the therapeutic response to transarterial chemoembolization is high, or a good prognosis is predicted, or

[0011] Step (b) above, in which, compared to the control group, the content of one or more bacteria of the family selected from the group consisting of Lachnospiraceae and Bacteroidaceae, bacteria of the genus Bacteroides, or bacteria of the species Ruminococcus gnavus is high, predicting a low therapeutic response to transarterial chemoembolization or a poor prognosis.

[0012] Another object of the present invention is to provide a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0013] (a) Step of separating control group and subject samples;

[0014] (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from the group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0015] (c) In the case where, compared to the control group in step (b) above, one or more alpha diversity indices selected from the group consisting of high ACE (abundance-based coverage estimators) indices, high CHAO 1 indices, and high Shannon indices,

[0016] A stage where the therapeutic response to transarterial chemoembolization is high or the prognosis is predicted to be good.

[0017] Another object of the present invention is to provide a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0018] (S1) Step of separating subject samples before and after carotid artery chemoembolization;

[0019] (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis; and

[0020] (S3) In the above step (S2), if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good, or

[0021] Step (S2) above, if the content of Ruminococcus gnavus species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is low or the prognosis is poor.

[0022] Another object of the present invention is to provide a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0023] (S1) Step of separating control and subject samples after carotid artery chemoembolization;

[0024] (S2) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0025] (S3) A step in which, compared to the control group in step (S2), one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0026]

[0027] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art to which the present invention belongs from the description below.

[0028] To achieve the above objectives, the present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0029] (a) Step of separating control group and subject samples;

[0030] (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis; and

[0031] (c) If, compared to the control group in step (b) above, the content of one or more bacteria of the family Prevotellaceae and Ruminococcaceae selected from the group consisting of Prevotella and Faecalibacterium, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, the therapeutic response to transarterial chemoembolization is high, or a good prognosis is predicted, or

[0032] Step (b) above, in which, compared to the control group, the content of one or more bacteria of the family selected from the group consisting of Lachnospiraceae and Bacteroidaceae, bacteria of the genus Bacteroides, or bacteria of the species Ruminococcus gnavus is high, predicting a low therapeutic response to transarterial chemoembolization or a poor prognosis.

[0033] In one embodiment of the present invention, the sample in step (a) may be a stool, but is not limited thereto.

[0034] In another embodiment of the present invention, in step (b), the metagenomic analysis may be performed by performing PCR (polymerase chain reaction) on the extracted DNA using the primer pair of SEQ ID NO. 1 and SEQ ID NO. 2, but is not limited thereto.

[0035] In another embodiment of the present invention, in step (b), the metagenomic analysis may analyze the microorganisms of the sample using 16S rRNA sequencing, but is not limited thereto.

[0036] In another embodiment of the present invention, the information providing method may analyze microorganisms of a sample at the species level using LEfSe (linear discriminant analysis Effect Size) analysis, but is not limited thereto.

[0037] In another embodiment of the present invention, based on the linear discriminant analysis (LDA) score of the LEfSe analysis, if one or more species of bacteria selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s are dominant among the total microorganisms in the subject sample, it can be predicted that the treatment response is high or the prognosis is good, but is not limited thereto.

[0038] In another embodiment of the present invention, based on the LDA score of the LEfSe analysis, if the Ruminococcus gnavus species is the dominant species among the total microorganisms in the subject sample, it can be predicted that the treatment response is low or the prognosis is poor, but is not limited thereto.

[0039] In another embodiment of the present invention, based on the linear discriminant analysis (LDA) score of the LEfSe analysis, if one or more species of bacteria selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s are the dominant species among the total microorganisms in the subject sample, it can be predicted that the treatment response is high or the prognosis is good, but is not limited thereto.

[0040] In another embodiment of the present invention, based on the LDA score of the LEfSe analysis, if the Ruminococcus gnavus species is the dominant species among the total microorganisms in the subject sample, it can be predicted that the treatment response is low or the prognosis is poor, but is not limited thereto.

[0041] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0042] (a) Step of separating control group and subject samples;

[0043] (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from the group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0044] (c) A step in which, compared to the control group in step (b) above, one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0045] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0046] (S1) Step of separating subject samples before and after carotid artery chemoembolization;

[0047] (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis; and

[0048] (S3) In the above step (S2), if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good, or

[0049] Step (S2) above, if the content of Ruminococcus gnavus species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is low or the prognosis is poor.

[0050] In one embodiment of the present invention, the sample in step (S1) may be a stool sample, but is not limited thereto.

[0051] In another embodiment of the present invention, in step (S2), the metagenomic analysis may be performed by using the primer pair of SEQ ID NO. 1 and SEQ ID NO. 2 on the extracted DNA, but is not limited thereto.

[0052] In another embodiment of the present invention, in step (S2), the metagenomic analysis may analyze the microorganisms of the sample using 16S rRNA sequencing, but is not limited thereto.

[0053] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0054] (S1) Step of separating control and subject samples after carotid artery chemoembolization;

[0055] (S2) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0056] (S3) A step in which, compared to the control group in step (S2), one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0057]

[0058] In addition, the present invention provides a method for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0059] (a) Step of separating control group and subject samples;

[0060] (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis; and

[0061] (c) If, compared to the control group in step (b) above, the content of one or more bacteria of the family Prevotellaceae and Ruminococcaceae selected from the group consisting of Prevotella and Faecalibacterium, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, the therapeutic response to transarterial chemoembolization is high, or a good prognosis is predicted, or

[0062] Step (b) above, in which, compared to the control group, the content of one or more bacteria of the family selected from the group consisting of Lachnospiraceae and Bacteroidaceae, bacteria of the genus Bacteroides, or bacteria of the species Ruminococcus gnavus is high, predicting a low therapeutic response to transarterial chemoembolization or a poor prognosis.

[0063] In addition, the present invention provides a method for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps:

[0064] (a) Step of separating control group and subject samples;

[0065] (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from the group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0066] (c) A step in which, compared to the control group in step (b) above, one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0067] In addition, the present invention provides a method for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps:

[0068] (S1) Step of separating subject samples before and after carotid artery chemoembolization;

[0069] (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis; and

[0070] (S3) In the above step (S2), if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good, or

[0071] Step (S2) above, if the content of Ruminococcus gnavus species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is low or the prognosis is poor.

[0072] In addition, the present invention provides a method for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps:

[0073] (S1) Step of separating control and subject samples after carotid artery chemoembolization;

[0074] (S2) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0075] (S3) A step in which, compared to the control group in step (S2), one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0076] In addition, the present invention relates to a transarterial chemoembolization composition comprising, as an active ingredient, a preparation for measuring the content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for predicting therapeutic responsiveness or prognosis for chemoembolization (TACE).

[0077] In addition, the present invention relates to transarterial chemoembolization of a preparation for measuring the content of one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for predicting treatment responsiveness or prognosis for TACE.

[0078] In addition, the present invention relates to a transarterial chemoembolization composition comprising, as an active ingredient, a preparation for measuring the content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for manufacturing a preparation for predicting therapeutic responsiveness or prognosis for chemoembolization (TACE).

[0079] In addition, the present invention relates to a preparation for the manufacture of a preparation for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or a selection from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus Provides a use for a preparation for measuring the content of one or more species of bacteria.

[0080] The intestinal microbiota of the present invention serves as a biomarker for predicting the therapeutic responsiveness or prognosis of transarterial chemoembolization in liver cancer patients. The intestinal microbiota of the present invention can be utilized as a potential predictor of clinical outcomes for transarterial chemoembolization, as an adjuvant therapeutic target for the treatment of liver cancer, and for predicting the survival rate of liver cancer patients who have undergone transarterial chemoembolization. Therefore, the biomarker of the present invention can be usefully utilized as a marker for predicting the therapeutic responsiveness or prognosis of transarterial chemoembolization in liver cancer patients.

[0081] Figure 1 shows the research flowchart.

[0082] Figures 2a and 2b show the reference characteristics of the entire cohort.

[0083] Figure 3 shows the initial response to transarterial chemoembolization (CR: complete remission, PR: partial response, SD: stable disease, PD: progressive disease).

[0084] Figures 4a and 4b show the baseline diversity and microbial composition of the gut microbiome of the responder group and the non-responder group, specifically Figure 4a shows the baseline alpha diversity of the gut microbiome of the responder group and the non-responder group, and Figure 4b shows the baseline beta diversity of the gut microbiome of the responder group and the non-responder group.

[0085] FIGS. 5a to 5c show baseline diversity and microbial composition of the gut microbiome of responder and non-responder groups. Specifically, FIG. 5a shows baseline microbial composition at the phylum level of responder and non-responder groups, FIG. 5b shows baseline microbial composition at the genus level of responder and non-responder groups, and FIG. 5c shows baseline microbial composition at the genus level of responder and non-responder groups.

[0086] Figures 5d and 5e show the LEfSe phylogenetic tree and reference gut microbiome plots for responder and non-responder groups; specifically, Figure 5d shows the LEfSe phylogenetic tree, and Figure 5e shows the LEfSe plot (LDA≥3). (LEfSe, linear discriminant analysis effect size; LDA, linear discriminant analysis)

[0087] Figures 5f and 5g show the relative abundance of responder-dominant and non-responder-dominant species in reference gut microbial communities of responder and non-responder groups, specifically Figure 5f shows responder-dominant species and Figure 5g shows non-responder-dominant species.

[0088] Figures 6a and 6b show the temporal change in alpha diversity of the gut microbial community in responder and non-responder groups, specifically Figure 6a shows the responder group and Figure 6b shows the non-responder group.

[0089] Figures 6c and 6d show the temporal changes in gut microbial community beta diversity of responder and non-responder groups, specifically Figure 6c shows the responder group and Figure 6d shows the non-responder group.

[0090] Figures 7a and 7b show the temporal changes in the relative abundance of Rubeola cecicola, Dialister_uc, PA001046_g_uc, DQ797020_s2 and Ruminococcus gnavus in the stool of the responder group and the non-responder group, specifically Figure 7a shows the responder group and Figure 7b shows the non-responder group.

[0091] Figures 8a and 8b show the overall survival rate and progression-free survival rate of all patients; specifically, Figure 8a shows the overall survival rate, and Figure 8b shows the progression-free survival rate.

[0092] Figures 9a to 9d show the overall survival rate and progression-free survival rate according to the abundance of Roseburia cecicola and Dialister_uc. Specifically, Figure 9a shows the overall survival rate according to the abundance of Roseburia cecicola, Figure 9b shows the overall survival rate according to the abundance of Dialister_uc, Figure 9c shows the progression-free survival rate according to the abundance of Roseburia cecicola, and Figure 9d shows the progression-free survival rate according to the abundance of Dialister_uc.

[0093] Figures 9e and 9f show the baseline characteristics of patients based on low and high abundance subgroups of Roseburia cecicola and Dialister_uc.

[0094] Figures 9g and 9h show the overall survival rate and progression-free survival rate according to the abundance of Ruminococcus gnavus. Specifically, Figure 9g shows the overall survival rate according to the abundance of Ruminococcus gnavus, and Figure 9h shows the progression-free survival rate according to the abundance of Ruminococcus gnavus.

[0095] The human gut microbiome plays a key role in the homeostasis between the gut and the liver, a relationship defined as the "Gut-Liver axis." In this invention, changes in specific gut microbial components have been proposed as potential biomarkers for the early diagnosis and prediction of the response to immunotherapy in hepatocellular carcinoma (HCC) and other solid tumors. Transarterial chemoembolization (TACE), a standard treatment for intermediate-stage HCC patients and a treatment option for advanced HCC patients, can displace bacteria by disrupting the intestinal barrier and temporarily increasing portal pressure. However, there have been no studies on the association between the gut microbiome and the clinical outcomes of TACE. Therefore, the inventors aimed to prospectively evaluate the association between the gut microbiome and survival outcomes and the response to TACE in patients with HCC.

[0096] The inventors presented a single-center prospective cohort study involving 96 hepatocellular carcinoma patients who underwent transarterial chemoembolization (TACE) at Asan Medical Center, a high-dose liver cancer center in Korea, between April 2021 and November 2023. Stool samples were collected at three time points: immediately before TACE (P0), one day after TACE (P1), and one month after TACE (P2). Stool 16S rRNA sequencing was analyzed to evaluate gut microbial diversity, composition, and dynamic changes. According to the mRECIST (modified Response Evaluation Criteria in Solid Tumors) criteria, 63 participants (65.6%) were early responders and 33 (34.4%) were early non-responders. The gut microbial community of early responders had higher alpha diversity and was richer in short-chain fatty acid (SCFA)-producing bacteria compared to early non-responders at baseline. The alpha-diversity of the microbial communities of both groups decreased immediately after transarterial chemoembolization and recovered within one month, but early non-responders showed more distinct changes in microbial composition over time compared to the microbial communities of early responders.

[0097] The inventors have identified an association between microbial epidemiology and clinical outcomes in hepatocellular carcinoma patients treated with transarterial chemoembolization. These results suggest that baseline microbial diversity and specific microbial components can be used as potential predictors of clinical outcomes and as adjuvant therapeutic targets for these patients.

[0098]

[0099] Including the entire claims below, the present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps:

[0100] (a) Step of separating control group and subject samples;

[0101] (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis; and

[0102] (c) If, compared to the control group in step (b) above, the content of one or more bacteria of the family Prevotellaceae and Ruminococcaceae selected from the group consisting of Prevotella and Faecalibacterium, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, the therapeutic response to transarterial chemoembolization is high, or a good prognosis is predicted, or

[0103] Step (b) above, in which, compared to the control group, the content of one or more bacteria of the family selected from the group consisting of Lachnospiraceae and Bacteroidaceae, bacteria of the genus Bacteroides, or bacteria of the species Ruminococcus gnavus is high, predicting a low therapeutic response to transarterial chemoembolization or a poor prognosis.

[0104] Including the entire claims below, the present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps:

[0105] (a) Step of separating control group and subject samples;

[0106] (b) a step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Roseburia cecicola and Dialister_uc through metagenome analysis; and

[0107] (c) A step in which, compared to the control group in step (b) above, the content of one or more species of bacteria selected from the group consisting of Roseburia cecicola and Dialister_uc is high, and the therapeutic responsiveness to transarterial chemoembolization is predicted to be high or the prognosis to be good.

[0108] Including all claims below, changes in the composition of the gut microbiome during hepatectomy as a curative treatment for liver cancer and hepatocellular carcinoma may be associated with postoperative liver degeneration and liver-related morbidity, which may have been confirmed through clinical and preclinical studies, but are not limited thereto.

[0109] Including all claims below, transarterial chemoembolization as a palliative treatment may be a treatment option for patients with a wide spectrum of liver cancer and hepatocellular carcinoma, but is not limited thereto.

[0110] Including all claims below, the present specification has a problem in that transarterial chemoembolization can have a transient effect on hepatic hemodynamics by reducing hepatic arterial blood flow and increasing portal blood pressure, and this physiological mechanism can lead to intestinal wall edema and intestinal wall dysfunction, which can cause the intestinal-hepatic axis to collapse. In addition, while microbial imbalance resulting from the collapse of the intestinal-hepatic axis can increase the risk of hepatocellular carcinoma, there has been a technical limitation in that there have been no clinical studies on the association between the intestinal microbiome and the therapeutic prognosis of transarterial chemoembolization in hepatocellular carcinoma patients.

[0111] Given that, as per the entire claim below, combination therapy including transarterial chemoembolization and immunotherapy or targeted molecular agents has shown better clinical outcomes than systemic monotherapy in unresectable hepatocellular carcinoma, positive results can be derived that additionally controlling bacteria related to the therapeutic responsiveness or prognostic prediction of the transarterial chemoembolization of the present invention can further improve treatment outcomes, but is not limited thereto.

[0112] In all claims below, the “gut microbiome” is proposed as a potential immunomodulator in various tumor microenvironments (TMEs) and may contribute to antitumor effects and tumor progression, but is not limited thereto. Furthermore, in the present invention, the gut microbiome may play a key role in intestinal inflammation and chronic inflammatory liver disease, and disturbance of the gut microbiome may induce bacterial metastasis and chronic inflammation of the liver, ultimately leading to liver fibrosis and hepatocellular carcinoma. Therefore, as identified in the present invention, specific changes in the gut microbiome may be considered as potential biomarkers for the early diagnosis and treatment response of hepatocellular carcinoma, but are not limited thereto.

[0113] In this specification, including the full claims below, “transarterial chemoembolization (TACE)” may be a method that increases drug delivery to a tumor with less systemic toxicity while having the effects of conventional embolization and cytotoxic chemotherapy. Portal pressure may temporarily increase during chemoembolization, which can cause the portal vein to divert intestinal blood to the liver, leading to bacterial metastasis and liver inflammation. In other words, transarterial chemoembolization can increase portal pressure and induce dysbiosis. However, there was a limitation in that there was no prior art regarding the association between gut microbiota and the clinical outcomes of transarterial chemoembolization. Accordingly, the inventors disclosed an invention that confirms the association between gut microbiota and clinical outcomes in patients with liver cancer or hepatocellular carcinoma treated with transarterial chemoembolization.

[0114] Including all claims below, in this specification, for transarterial chemoembolization, one or more selected from the group consisting of doxorubicin, mitomycin, and cisplatin may be injected through an artery, specifically, one or more selected from the group consisting of doxorubicin and cisplatin may be injected through an artery, but is not limited thereto. The artery may be an artery that supplies blood to liver cancer or hepatocellular carcinoma.

[0115] Including all claims below, the “method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization” may be a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization in liver cancer patients, but is not limited thereto.

[0116] In the entire specification including the following claims, the prediction of “treatment responsiveness” may include, but is not limited to, predicting whether the prognosis will be good or bad when transarterial chemoembolization is performed.

[0117] Including all claims below, the term “prognosis prediction” in this specification may mean, but is not limited to, the prognosis prediction of a patient with liver cancer or hepatocellular carcinoma.

[0118] In this specification, including all claims below, “forecast” may be used interchangeably with “risk forecast” and “risk forecast,” but is not limited thereto.

[0119] In all claims below, “therapeutic responsiveness or prognostic prediction” may be determined by whether a patient has achieved one or more of the groups consisting of complete remission (CR), partial remission (PR), stable disease (SD), and progressive disease (PD) using the modified Response Evaluation Criteria in Solid Tumors (mRECIST), but is not limited thereto. Additionally, it may refer to an association between survival outcomes such as overall survival (OS) and progression-free survival (PFS), but is not limited thereto.

[0120] In the entire specification including the following claims, “treatment responsiveness or prognosis prediction” may mean an association between transarterial chemoembolization and survival outcomes, but is not limited thereto; specifically, in the present invention, “an association between transarterial chemoembolization and survival outcomes” refers to performing transarterial chemoembolization and 1 to 40 months, 1 to 35 months, 1 to 30 months, 1 to 25 months, 1 to 20 months, 1 to 15 months, 1 to 10 months, 1 to 5 months, 5 to 40 months, 5 to 35 months, 5 to 30 months, 5 to 25 months, 5 to 20 months, 5 to 15 months, 5 to 10 months, 10 to 40 months, 10 to 35 months, 10 to 30 months, 10 to 25 months, 10 to 20 months, 10 to 15 months, 15 to 40 months, 15 to 35 months, 15 to 30 months, 15 to 25 months, 15 to 20 months, 20 to 40 months, 20 to 35 months, 20 to 30 months, 20 to 25 months, 25 to 40 months, 25 to 35 months, 25 to 30 months, 30 to 40 months, 30 to 35 months, 35 to 40 months, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, It may imply an association with survival outcomes after 25, 26, 27, 28, 29, or 30 months, but is not limited thereto.

[0121] In the entirety of the following claims, the term “prediction of treatment responsiveness or prognosis” may be used interchangeably with “determining whether there is susceptibility or resistance to treatment” and “method for providing information for diagnosing resistance,” but is not limited thereto. In the present invention, “predicting low treatment responsiveness or poor prognosis” may be used interchangeably with “determining a patient to be resistant to treatment,” but is not limited thereto.

[0122] In the entirety of the following claims, “predicted to have a low therapeutic response to transarterial chemoembolization (TACE) or a poor prognosis” may mean “determined to have a low therapeutic response to transarterial chemoembolization or a high risk of liver cancer progression”; “determined to have a low therapeutic response to transarterial chemoembolization or a high risk of liver cancer development”; “determined to have a low therapeutic response to transarterial chemoembolization or a high risk of liver cancer aggravation”; and “determined to have a low therapeutic response to transarterial chemoembolization or a high risk of liver cancer metastasis”, but is not limited thereto. A person skilled in the art can understand the meaning of the opposite term as having the opposite meaning in accordance with the above definitions.

[0123] In the entirety of the following claims, the “control group” may be a person or other organism having liver cancer and / or hepatocellular carcinoma, and said organism may mean mammals such as non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0124] In all claims below, the term “subject” may be a person or other organism suspected of having liver cancer and / or hepatocellular carcinoma, and said organism may mean non-human mammals such as primates, mice, rats, dogs, cats, horses, and cattle. Additionally, the term “subject” may be used interchangeably with “patient,” but is not limited thereto.

[0125] In all claims below, the term “patient” in this specification refers to a subject requiring diagnosis of a disease, treatment responsiveness, or prognosis prediction, and more specifically refers to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle. Additionally, in the present invention, the term “patient” may refer to a patient with high or low treatment responsiveness to transarterial chemoembolization, or a responder or non-responder to transarterial chemoembolization, but is not limited thereto. Furthermore, the term “patient” may be used interchangeably with “individual,” but is not limited thereto.

[0126] In this specification, including all claims below, liver cancer means any type of malignant tumor occurring in the liver and may be hepatocellular carcinoma (HCC), or one or more cancers selected from the group consisting of hepatoblastoma, cholangiocarcinoma, cholangiocarcinoma cystadenocarcinoma, or liver cancer arising from viral infection. Additionally, hepatocellular carcinoma may correspond to the most major histological subtype, accounting for 70 to 85% of primary liver cancers, but is not limited thereto.

[0127] In this specification, including all claims below, the term "metagenome" refers to the sum of genomes including all viruses, bacteria, fungi, etc., within an isolated area, such as soil or the intestines of animals. It may be a concept of a genome that explains the identification of a large number of microorganisms at once using a sequencer to analyze microorganisms that cannot be cultured. Furthermore, in the present invention, the metagenome does not refer to the genome or genome of a single species, but rather to a type of mixed genome consisting of the genomes of all species within a single environmental unit. This term emerged during the process of biological development in an omics-oriented manner, from the perspective that when defining a species, it involves not only a single functionally existing species but also the interaction of various species to form a complete species. Technically, it may be the subject of a technique that uses rapid sequencing methods to analyze all DNA and RNA regardless of species, thereby identifying all species within an environment and elucidating their interactions and metabolic processes.

[0128] Including all claims below, metagenome analysis may be performed by performing PCR, using 16S rRNA sequencing, or using LEfSe analysis, but is not limited thereto.

[0129] Including all claims below, in this specification, one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus may be intestinal bacteria, but are not limited thereto. In addition, the above bacteria may be a biomarker of the present invention.

[0130] In the entirety of the following claims, “bacteria” may be intestinal bacteria, specifically, short-chain fatty acid-producing bacteria or bacteria that do not produce short-chain fatty acids, but are not limited thereto.

[0131] In all claims below, the term “bacteria” may be used interchangeably with “bacteria,” and in the present invention, the term “bacteria” is a concept included in, but not limited to, “microorganisms.”

[0132] In the entirety of the following claims, the term “Prevotellaceae” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) to be Prevotellaceae by a genomic analysis means known in the art.

[0133] In the entirety of the following claims, the term “Ruminococcaceae” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) to be Ruminococcaceae by a genomic analysis means known in the art.

[0134] In the entirety of the following claims, the term “Lachnospiraceae” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) to be Lachnospiraceae by a genomic analysis means known in the art.

[0135] In the entirety of the following claims, the term “Bacteroidaceae” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) to be Bacteroidaceae by a genomic analysis means known in the art.

[0136] In the entirety of the following claims, “Prevotella” is not limited to any specific type of bacterium, provided that it is known in the art or is determined (classified) as Prevotella by a genomic analysis means known in the art.

[0137] In the entirety of the following claims, “Faecalibacterium” is not limited to any specific type of bacterium, provided that it is a bacterium known in the art or determined (classified) to be Faecalibacterium by a genomic analysis means known in the art.

[0138] In the entirety of the following claims, “Bacteroides” is not limited to any specific type, provided it is a bacterium known in the art or determined (classified) as Bacteroides by a genomic analysis means known in the art.

[0139] In the entirety of the following claims, “Roseburia cecicola” is not limited to any specific type, provided it is a bacterium known in the art or determined (classified) as Roseburia cecicola by a genomic analysis means known in the art.

[0140] In the entirety of the following claims, “Dialister_uc” is not limited to any specific type, provided it is a bacterium known in the art or determined (classified) as Dialister_uc by a genomic analysis means known in the art.

[0141] In the entirety of the following claims, “PAC001046” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) as PAC001046 by a genomic analysis means known in the art.

[0142] In the entirety of the following claims, “DQ797020_s” is not limited to any specific type of bacteria, provided that it is a bacterium known in the art or determined (classified) as DQ797020_s by a genomic analysis means known in the art.

[0143] In the entire specification including the following claims, “Ruminococcus gnavus” is not limited to any type of bacterium, provided it is a bacterium known in the art or determined (classified) as Ruminococcus gnavus by a genomic analysis means known in the art.

[0144] In the entirety of the following claims, “decision” may be used interchangeably with determination, instruction, or meaning.

[0145] Including all claims below, the bacteria (microorganisms) of the present invention described above may be used in combination of two or more, but are not limited thereto.

[0146] Including the full claims below, the two groups (responders and non-responders) differed in the relative taxonomic abundance of specific gut microbial communities at baseline, and the responders were more abundant in the genera Prevotella and Faecalibacterium than the non-responders at baseline.

[0147] Including the entire claims below, microorganisms of the genera Prevotella and Faecalibacterium may be intestinal microorganisms and may be bacteria that produce short-chain fatty acids. Additionally, they may have a positive effect on the therapeutic response to anti-programmed death (PD)-1 and anti-PD-L1 (PD-Ligand 1, programmed death-ligand 1) immunotherapy in other solid tumors such as colorectal cancer (CRC), melanoma, renal cell carcinoma, and lung cancer, but are not limited thereto.

[0148] Including all claims below, the short-chain fatty acid-producing bacteria Dialister_uc and Roseburia cecicola may be abundant in the microbial communities of patients with hepatocellular carcinoma treated with immunotherapy, but are not limited thereto.

[0149] Including all claims below, the abundance of Dialister_uc and Roseburia cecicola at baseline may be used as a prognostic indicator to predict treatment response and survival outcomes after transarterial chemoembolization, and particularly in the case of Dialister_uc, survival outcomes of TACE may be associated independently of patient and tumor characteristics, but are not limited thereto.

[0150] In the entire specification including the following claims, “bacterial content” may mean “abundance,” and “abundance” may mean relative abundance, but is not limited thereto. Additionally, in the present invention, bacterial content may mean one or more contents selected from the group consisting of phylum level, family level, genus level, and species level, but is not limited thereto.

[0151] Additionally, in all claims below, the term “content of species bacteria” may mean relative abundance at the species level and may mean calculated as the ratio of the number of OTUs (Operational Taxonomic Units) belonging to each species in a phylogenetic tree, but is not limited thereto.

[0152] In the entire specification including the following claims, the content of the bacteria of the present invention was measured once or two or more times over time, and the content was analyzed for therapeutic responsiveness and prognosis to transarterial chemoembolization, and it was confirmed that there is a significant correlation.

[0153] In the entirety of the following claims, the term "baseline" may refer to the first point in time of the timeline for measuring the bacterial content of the present invention, and may mean the most reference point in a series of bacterial content measurements. Additionally, the term "reference level" may refer to the bacterial content at the "baseline."

[0154] Including the entire claims below, the “comparison” refers to one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus, obtained by extracting DNA from samples isolated from the same or different patients and analyzing them through metagenomic analysis. It may involve comparing bacterial content, specifically, through metagenomic analysis of DNA extracted from samples isolated from the same or different patients, regarding the content of one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus It may involve comparing each one, but is not limited to this.Furthermore, it is a comparison made using any method commonly used in the industry, and it is a broad concept that includes both quantitative and qualitative comparisons.

[0155] Including all claims below, the “content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus” comprises four bacteria of families and three genera It may mean the content of one or more bacteria selected from a group consisting of (genus) bacteria and five species (species) bacteria, but is not limited thereto.

[0156] In the entirety of the following claims, the term “high content” may be used interchangeably with “increased content.” In this case, “high content” means that something previously undetectable is detected, or that the amount detected is relatively higher than the normal content. For example, “increased content” means that the bacterial content in the sample is at least 1%, 2%, 3%, 4%, 5%, 10% or higher, e.g., 5%, 10%, 20%, 30%, 40%, or 50%, 60%, 70%, 80%, 90% or higher, and / or 0.5 times, 1.1 times, 1.2 times, 1.4 times, 1.6 times, 1.8 times or higher. Specifically, it may mean that compared to that of the control group, it is increased by 1 to 1.5 times, 1.5 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 3.5 times, 3.5 to 4 times, 4 to 4.5 times, 4.5 to 5 times, 5 to 5.5 times, 5.5 to 6 times, 6 to 6.5 times, 6.5 to 7 times, 7 to 7.5 times, 7.5 to 8 times, 8 to 8.5 times, 8.5 to 9 times, 9 to 9.5 times, 9.5 to 10 times, or 10 times or more, but is not limited thereto. Furthermore, in the present invention, “high bacterial content” may have a broad meaning including a statistically significant level, but is not limited thereto. To a person skilled in the art, the meaning of the term opposite to this can be understood as having the opposite meaning in accordance with the above definition.

[0157] In this specification, including the entire claims below, “analysis” may be used interchangeably with “identification,” “measurement,” or “detection” (and conversely interchangeably), and may include quantifying the content, etc., of a detected or measured object; and since it includes a qualitative meaning of confirming the presence or absence of a specific substance, it includes both measuring and confirming the presence (expression) of a target substance and measuring and confirming a change in the presence level (expression level) of a target substance.

[0158] In the entirety of the following claims, “sample” may be one or more selected from the group consisting of blood, serum, whole blood, plasma, urine, feces, saliva, tissue, cell, organ, bone marrow, fine needle aspiration specimen, core needle biopsy specimen, and vacuum aspiration biopsy specimen, but is not limited thereto.

[0159] Additionally, throughout the following claims, the term "sample" may be used interchangeably with biological samples and biological samples isolated from a patient, but is not limited thereto.

[0160] In all claims below, the “biological sample” in this specification may be a biological sample isolated from a patient with liver cancer or hepatocellular carcinoma, but is not limited thereto. Additionally, in the present invention, the “isolated biological sample” may be isolated before, after, or regardless of the course of transarterial chemoembolization treatment, but is not limited thereto.

[0161] Including all claims below, in step (a) of the present invention, the sample may be a stool, but is not limited thereto.

[0162] Including all claims below, in the specification, the metagenome analysis in step (b) of the present invention may be performed by performing PCR (polymerase chain reaction) on the extracted DNA of the present invention using the primer pair of SEQ ID NO. 1 and SEQ ID NO. 2, but is not limited thereto.

[0163] Including the entire claims below, in step (b) of the present invention, metagenome analysis may be performed by analyzing the microorganisms of the sample of the present invention using 16S rRNA sequencing, but is not limited thereto.

[0164] Including the full claims below, in step (b), after 16S rRNA sequencing of the sample of the present invention, the bacteria of the sample may be classified through OTU (Operational taxonomic unit) analysis, but are not limited thereto.

[0165] Including all claims below, the information-providing method of the present invention may analyze microorganisms of a sample at the species level using LEfSe (linear discriminant analysis (LDA) Effect Size) analysis, but is not limited thereto.

[0166] Including the entire claims below, in step (b) of the invention, the metagenome analysis may be the analysis of microorganisms in a sample at the species level using LEfSe (linear discriminant analysis (LDA) Effect Size) analysis, but is not limited thereto.

[0167] Including the full claims below, if, based on the LDA score, one or more species of bacteria selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s are dominant in the subject sample, it can be predicted that there is a high therapeutic response or a good prognosis, but is not limited thereto.

[0168] Including all claims below, the method for providing information of the present invention may further include, but is not limited to, a step of predicting that the treatment responsiveness is high or the prognosis is good when, based on the LDA score, one or more species of bacteria selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s are dominant species among the total microorganisms in a subject sample.

[0169] Including all claims below, if Ruminococcus gnavus species is the dominant species among the total microorganisms in a subject sample based on the LDA score, it can be predicted that the treatment response will be low or the prognosis will be poor, but is not limited thereto.

[0170] Including all claims below, the information-providing method of the present invention may further include, but is not limited to, a step of predicting that the treatment responsiveness is low or the prognosis is poor when Ruminococcus gnavus species is the dominant species among all microorganisms in a subject sample based on the LDA score.

[0171] Including all claims below, LEfSe may be, but is not limited to, an algorithm for the discovery and description of high-dimensional biomarkers that identifies genomic features (genes, pathways, or taxa) characterizing differences between two or more biological conditions (or classes).

[0172] Including all claims below, as provided in this specification, LDA may be used in LEfSe to estimate the effect size of each differentially rich feature, but is not limited thereto.

[0173] Including all claims below, in this specification, analyzing microorganisms of a sample at the species level may be analyzing microorganisms based on LDA scores, but is not limited thereto.

[0174] In the entire specification including the following claims, “when species bacteria are the dominant species” may mean that the LDA score (Segata et al. Genome Biology 2011, 12:R60) is 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 or higher, and specifically may mean that the LDA score is 2, 2.5, 3, 3.5, or 4 or higher, but is not limited thereto.

[0175] In the entire specification including the following claims, “when the species of bacteria is the dominant species” may mean that the LDA score is 3 or higher, but is not limited thereto. Specifically, in one embodiment of the present invention, if the LDA score is higher than 3, the corresponding variable (specifically, microorganisms or bacteria) may not be derived. Furthermore, if the LDA score is lower than 3, the corresponding variable may not only fail to reflect the difference between the subject and the control group, but the variable may also be derived excessively. Therefore, determining that the LDA score is 3 or higher may be a suitable criterion for determining “when the species of bacteria is the dominant species,” but is not limited thereto.

[0176] In this specification, including the entire claims below, "dominant species" and "predominant species" may be used interchangeably.

[0177] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0178] (a) Step of separating control group and subject samples;

[0179] (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from the group consisting of the ACE (Abundance-based Coverage Estimator) index, the CHAO 1 index, and the Shannon index through metagenome analysis; and

[0180] (c) A step in which, compared to the control group in step (b) above, one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0181] Including all claims below, the information providing method may be applied without limitation where the above-described content can be applied.

[0182] Including all claims below, the information providing method described in the present invention may be characterized by further including the steps of the information providing method, but is not limited thereto.

[0183] In all claims below, the “alpha diversity index” is for measuring bacterial diversity within a microbial community and may be an alpha diversity index regarding intestinal bacterial diversity, and may be, for example, an alpha diversity index regarding short-chain fatty acid-producing bacteria, but is not limited thereto.

[0184] In the entirety of the claims below, “alpha diversity index” means diversity within a single sample, and “beta diversity index” may mean variation in species composition and richness when comparing multiple samples or groups, but is not limited thereto.

[0185] Including the full claims below, the present invention compared baseline alpha-diversity between responders and non-responders and was able to confirm that responders exhibited higher alpha-diversity. Additionally, for hepatocellular carcinoma patients who received immunotherapy, responders may exhibit a higher alpha-diversity Shannon index than non-responders, which may mean that they exhibited greater species abundance at baseline.

[0186] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0187] (S1) Step of separating subject samples before and after carotid artery chemoembolization;

[0188] (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis; and

[0189] (S3) In the above step (S2), if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good, or

[0190] Step (S2) above, if the content of Ruminococcus gnavus species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is low or the prognosis is poor.

[0191] Including all claims below, the information providing method may be applied without limitation where the above-described content can be applied.

[0192] Including all claims below, the information providing method described in the present invention may be characterized by further including the steps of the information providing method, but is not limited thereto.

[0193] In the entire specification including the following claims, “before transarterial chemoembolization” may be 1 to 10 days, 1 to 9 days, 1 to 8 days, 1 to 7 days, 1 to 6 days, 1 to 5 days, 1 to 4 days, 1 to 3 days, or 1 to 2 days before transarterial chemoembolization, and specifically may be 3 days, 2 days, or 1 day before, but is not limited thereto.

[0194] Including all claims below, in this specification, “after transarterial chemoembolization” refers to days 1 to 60 days, 1 to 55 days, 1 to 50 days, 1 to 45 days, 1 to 40 days, 1 to 35 days, 1 to 30 days, 5 to 60 days, 5 to 55 days, 5 to 50 days, 5 to 45 days, 5 to 40 days, 5 to 35 days, 5 to 30 days, 10 to 60 days, 10 to 55 days, 10 to 50 days, 10 to 45 days, 10 to 40 days, 10 to 35 days, 10 to 30 days, 15 to 60 days, 15 to 55 days, 15 days to 50 days, 15 to 45 days, 15 to 40 days, 15 to 35 days, 15 to 30 days, 20 to 60 days, 20 to 55 days, 20 to 50 days, 20 to 45 days, 20 to 40 days, 20 to 35 days, 20 to 30 days, 25 to 60 days, 25 to 55 days, 25 to 50 days, 25 to 45 days, 25 to 40 days, 25 to 35 days, 25 to 30 days, 30 to 60 days, 30 to 55 days, 30 to 50 days, 30 to 45 days, 30 to 40 days, or 30 to 35 days It may be later, or on the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 41st, 42nd, 43rd, 44th, 45th, 46th, 47th, 48th, 49th, It may be 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days later, but is not limited thereto.

[0195] In the entirety of the following claims, the term “high content” may be used interchangeably with “increased content.” In this case, “high content” means that something previously undetectable is detected, or that the amount detected is relatively higher than the normal content. For example, “increased content” means that the bacterial content in a sample is at least 1%, 2%, 3%, 4%, 5%, 10% or higher, e.g., 5%, 10%, 20%, 30%, 40%, or 50%, 60%, 70%, 80%, 90% or higher, and / or 0.5 times, 1.1 times, 1.2 times, 1.4 times, 1.6 times, 1.8 times or higher. Specifically, it may mean that compared to that of another period, it has increased by 1 to 1.5 times, 1.5 to 2 times, 2 to 2.5 times, 2.5 to 3 times, 3 to 3.5 times, 3.5 to 4 times, 4 to 4.5 times, 4.5 to 5 times, 5 to 5.5 times, 5.5 to 6 times, 6 to 6.5 times, 6.5 to 7 times, 7 to 7.5 times, 7.5 to 8 times, 8 to 8.5 times, 8.5 to 9 times, 9 to 9.5 times, 9.5 to 10 times, or more than 10 times, but is not limited thereto. Furthermore, in the present invention, “high bacterial content” may have a broad meaning including a statistically significant level, but is not limited thereto. To a person skilled in the art, the meaning of the term opposite to this can be understood as having the opposite meaning in accordance with the above definition.

[0196] Including all claims below, in step (S1) of the present invention, the sample may be a stool, but is not limited thereto.

[0197] Including all claims below, in step (S2) of the present invention, the metagenomic analysis may be performed by using the primer pair of SEQ ID NO. 1 and SEQ ID NO. 2 on the extracted DNA of the present invention, but is not limited thereto.

[0198] Including the entire claims below, in step (S2) of the present invention, metagenome analysis may be performed by analyzing the microorganisms of the sample of the present invention using 16S rRNA sequencing, but is not limited thereto.

[0199] The present invention provides a method for providing information for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization, comprising the following steps:

[0200] (S1) Step of separating control and subject samples after carotid artery chemoembolization;

[0201] (S2) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and

[0202] (S3) A step in which, compared to the control group in step (S2), one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis.

[0203] Including all claims below, the information providing method may be applied without limitation where the above-described content can be applied.

[0204] Including all claims below, the information providing method described in the present invention may be characterized by further including the steps of the information providing method, but is not limited thereto.

[0205]

[0206] The present invention provides a method for treating liver cancer comprising the following steps:

[0207] (a) Step of separating control group and subject samples;

[0208] (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis;

[0209] (c) a step in which, compared to the control group in step (b) above, if the content of one or more bacteria of the family selected from the group consisting of Prevotellaceae and Ruminococcaceae, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, it is predicted that the therapeutic response to transarterial chemoembolization is high or the prognosis is good; and

[0210] (d) A step of treating by performing transarterial chemoembolization when the therapeutic response to the above transarterial chemoembolization is high or when a good prognosis is predicted.

[0211] In addition, the present invention provides a method for treating liver cancer comprising the following steps:

[0212] (a) Step of separating control group and subject samples;

[0213] (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis;

[0214] (c) a step of predicting a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis when having one or more alpha diversity indices selected from a group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index compared to the control group in step (b) above; and

[0215] (d) A step of treating by performing transarterial chemoembolization when the therapeutic response to the above transarterial chemoembolization is high or when a good prognosis is predicted.

[0216] In addition, the present invention provides a method for treating liver cancer comprising the following steps:

[0217] (S1) Step of separating subject samples before and after carotid artery chemoembolization;

[0218] (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis;

[0219] (S3) A step of predicting that if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization in the above step (S2), the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good; and

[0220] (S4) A step of performing transarterial chemoembolization to treat when the therapeutic response to the transarterial chemoembolization is high or when a good prognosis is predicted.

[0221] In addition, the present invention provides a method for treating liver cancer comprising the following steps:

[0222] (S1) Step of separating control and subject samples after carotid artery chemoembolization;

[0223] (S2) A step of extracting DNA from the above sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis;

[0224] (S3) A step in which, in the above step (S2), one or more alpha diversity indices selected from a group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index compared to the control group are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis; and

[0225] (S4) A step of performing transarterial chemoembolization to treat when the therapeutic response to the transarterial chemoembolization is high or when a good prognosis is predicted.

[0226] In the entirety of the following claims, the phrase “the step of treating by performing transarterial chemoembolization when the therapeutic responsiveness to the transarterial chemoembolization is high or the prognosis is predicted to be good” may be used interchangeably with, but is not limited to, the phrase “the step of treating by performing transarterial chemoembolization on a subject when the therapeutic responsiveness to the transarterial chemoembolization is high or the prognosis is predicted to be good.”

[0227] Including all claims below, the liver cancer may be hepatocellular carcinoma, but is not limited thereto.

[0228] Including all claims below, the method for treating liver cancer described above may be applied in the same way where applicable, but is not limited thereto.

[0229] Including all claims below, the “method for treating liver cancer” may be applied simultaneously or sequentially with a general treatment method for treating liver cancer, but is not limited thereto.

[0230] Including all claims below, the “method for treating liver cancer” may be prescribed together with a pharmaceutical composition for prevention or treatment of liver cancer.

[0231] Including all claims below, the method for treating liver cancer may be applied simultaneously or sequentially with one or more selected from the group consisting of doxorubicin, mitomycin, and cisplatin, but is not limited thereto.

[0232] Including all claims below, the pharmaceutical composition for prevention or treatment of the present invention may further comprise a suitable carrier, excipient, and diluent commonly used in the manufacture of pharmaceutical compositions. The excipient may be one or more selected from the group consisting of, for example, diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.

[0233] Including all claims below, the pharmaceutical composition of the present invention may be used by being formulated in the form of a powder, granule, sustained-release granule, enteric granule, liquid, ophthalmic, oleic, emulsion, suspension, ethanol, troche, fragrance, limonene admixture, tablet, sustained-release tablet, enteric-coated tablet, sublingual tablet, hard capsule, soft capsule, sustained-release capsule, enteric-coated capsule, pill, tincture, soft extract, dry extract, fluid extract, injection, capsule, irrigation solution, warning agent, lotion, paste, spray, inhalant, patch, sterile injectable solution, or aerosol, etc., according to conventional methods, and said external preparation may be used in the form of a cream, gel, patch, spray, ointment, warning agent, lotion, liniment, paste, or cataplasma. Can have.

[0234] Including all claims below, carriers, excipients, and diluents that may be included in the pharmaceutical composition of the present invention may include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0235] Including all claims below, the formulation is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

[0236] Including all claims below, the present specification comprises, as additives to the tablets, powders, granules, capsules, pills, and lozenges of the present invention, excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium monohydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, refined lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethylcellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, primogel, etc.; Gelatin, gum arabic, ethanol, agar powder, cellulose phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium casein, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch paste, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. may be used as binders, and hydroxypropylmethylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, Calcium carboxymethylcellulose, calcium citrate, sodium lauryl sulfate, anhydrous silica, 1-hydroxypropylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium aluminum silicate, D-sorbitol solution, hard anhydrous silica, etc. disintegrants;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium pods, kaolin, petroleum jelly, sodium stearate, cocoa paste, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silica, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and hard anhydrous silica may be used.

[0237] Including all claims below, as used in this specification, additives to the liquid formulation of the present invention may include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, monostearic acid sucroses, polyoxyethylene sorbitol fatty acid esters (tween esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, water ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, etc.

[0238] Including all claims below, the syrup formulation of the present invention may use a solution of white sugar, other sugars or sweeteners, etc., and, if necessary, flavorings, coloring agents, preservatives, stabilizers, suspending agents, emulsifiers, viscosity enhancers, etc.

[0239] Including all claims below, the emulsion of the present invention may use purified water, and, if necessary, may use emulsifiers, preservatives, stabilizers, fragrances, etc. In this specification, including all claims below, purified water may be used in the emulsion of the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc.

[0240] Including all claims below, the suspending agent of the present invention may use acacia, tragacanthus, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc., and, if necessary, surfactants, preservatives, stabilizers, coloring agents, and fragrances may be used.

[0241] Including all claims below, the injectable formulation of the present invention comprises solvents such as distilled water for injection, 0.9% sodium chloride injection solution, Ringer's injection solution, dextrose injection solution, dextrose + sodium chloride injection solution, PEG, lactated Ringer's injection solution, ethanol, propylene glycol, non-volatile oils—sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; and solubilizing agents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tween, nijungtinamide, hexamine, and dimethylacetamide; Buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptones, and gums; isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid, and sodium bisulfite acetone; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; It may include suspending agents such as sodium CMC, sodium alginate, Tween 80, and aluminum monostearate.

[0242] Including all claims below, the suppositories of the present invention comprise: cocoa gluten, lanolin, Witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, lanette wax, glycerol monostearate, Tween or Spandex, Imhausen, monollene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydroccote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Bases such as Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Masa-MF, Masupol, Masupol-15, Neosupostal-N, Paramount-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wekovi (W, R, S, M, Fs), and Tegestor triglyceride base (TG-95, MA, 57) may be used.

[0243] In the entirety of the following claims, solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. with the extract. In addition, lubricants such as magnesium styrate and talc are also used in addition to simple excipients.

[0244] In the present specification, including all claims below, liquid formulations for oral administration include suspensions, liquid formulations, emulsions, syrups, etc., and may include various excipients, such as humectants, sweeteners, flavorings, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0245] Including all claims below, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The “pharmaceutically effective amount” means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient’s disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0246] Including all claims below, the pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects, taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art to which the present invention pertains.

[0247] Including the entire claims below, the pharmaceutical composition of the present invention may be administered to a patient by various routes. All modes of administration may be anticipated, for example, by oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal (intradural) injection, sublingual administration, buccal mucosal administration, rectal insertion, vaginal insertion, ocular administration, ear administration, nasal administration, inhalation, spray through the mouth or nose, skin administration, transdermal administration, etc.

[0248] Including the full claims below, the pharmaceutical composition of the present invention is determined by the type of drug as the active ingredient, along with various relevant factors such as the disease to be treated, the route of administration, the age, gender, weight, and severity of the disease of the patient.

[0249] In the entirety of the following claims, the term “patient” means a subject requiring treatment for a disease, and more specifically means mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0250] Including all claims below, the term “administration” in this specification means providing a predetermined composition of the present invention to a patient by any appropriate method.

[0251] In all claims below, the term “prevention” means any act of suppressing or delaying the onset of a target disease, the term “treatment” means any act of improving or beneficially altering a target disease and associated metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and the term “improvement” means any act of reducing parameters related to a target disease, e.g., the severity of symptoms, by administering a composition according to the present invention.

[0252] In addition, the present invention relates to a transarterial chemoembolization composition comprising, as an active ingredient, a preparation for measuring the content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for predicting therapeutic responsiveness or prognosis for chemoembolization (TACE).

[0253] In addition, the present invention relates to transarterial chemoembolization of a preparation for measuring the content of one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for predicting treatment responsiveness or prognosis for TACE.

[0254] In addition, the present invention relates to a transarterial chemoembolization composition comprising, as an active ingredient, a preparation for measuring the content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. It provides a use for manufacturing a preparation for predicting therapeutic responsiveness or prognosis for chemoembolization (TACE).

[0255] In addition, the present invention relates to a preparation for the manufacture of a preparation for predicting therapeutic responsiveness or prognosis for transarterial chemoembolization (TACE), comprising one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or a selection from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus Provides a use for a preparation for measuring the content of one or more species of bacteria.

[0256]

[0257] In this specification, including the entire set of claims below, the terms used in the present invention have been selected to be as widely used as possible, taking into account their functions in the invention; however, these terms may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc. Additionally, in specific cases, terms have been arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in the present invention should be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0258] Throughout the specification, including the claims of the present invention, when a part is described as “comprising” a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components. Throughout the specification of the present invention, terms such as “about,” “substantially,” etc., are used to mean at or near the stated value when inherent manufacturing and material tolerances are presented in the said sense, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which precise or absolute values ​​are mentioned to aid in understanding the present invention.

[0259] Throughout the specification, including the claims of the present invention, the term “combination thereof” included in the Markush-format expressions means one or more mixtures or combinations selected from the group consisting of the components described in the Markush-format expressions, and means including one or more selected from the group consisting of said components.

[0260] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided merely to facilitate a better understanding of the invention, and the scope of the invention is not limited by the following embodiments.

[0261]

[0262] [Example]

[0263]

[0264] Research design and variables

[0265]

[0266] This study was a single-center prospective cohort study of hepatocellular carcinoma patients who underwent transarterial chemoembolization (TACE) at Asan Medical Center, a high-dose liver cancer center in Seoul, South Korea, between April 2021 and November 2023. Participants who met all of the following criteria were eligible for enrollment: (i) age 18 or older, (ii) diagnosis of hepatocellular carcinoma, (iii) TACE treatment on the initial enrollment date, and (iv) if the participant had a history of previous TACE, the interval between the last TACE date and the index date had to be at least 6 weeks to ensure the recovery of microbial diversity. Additionally, only participants who signed an informed consent form were enrolled. Participants who met any of the following criteria were deemed ineligible: (i) failure to collect a reference stool sample prior to TACE, (ii) receiving treatment other than TACE, (iii) presence of a malignant tumor, or (iv) withdrawal of consent (Fig. 1). The follow-up end date was May 31, 2024. The index date was defined as the start date of transarterial chemoembolization performed during each participant's enrollment period. A total of 96 participants were enrolled. Participant characteristics included demographic findings, complications, etiology of hepatocellular carcinoma, and liver function at baseline. Additionally, data were obtained on baseline tumor characteristics, including tumor markers, chemotherapy agents used for transarterial chemoembolization, Barcelona Clinic Liver Cancer (BCLC) stage, modified Union for International Cancer Control (mUICC) stage, and prior treatment prior to transarterial chemoembolization. Dietary habits, smoking history, alcohol consumption, use of analgesics, probiotics, multivitamins and herbal medicines, and bowel habits were collected via questionnaires.

[0267] This study was approved by the Institutional Review Board of Asan Medical Center [IRB No. 2021-0240] and was conducted in accordance with the Institutional Research Board's ethical standards and the latest version of the Declaration of Helsinki. All participants signed written informed consent upon enrollment. In addition, the guidelines for reporting observational studies in epidemiology were followed.

[0268]

[0269] Carotid artery chemoembolization

[0270]

[0271] All eligible patients underwent transarterial chemoembolization after sample collection prior to treatment. For transarterial chemoembolization, 50 mg of doxorubicin or 2 mg / kg of cisplatin was infused through the artery supplying blood to the hepatocellular carcinoma (HCC) following selective catheterization of the supplying artery (Clin Mol Hepatol. 7 2023;29(3):521-541).

[0272]

[0273] Stool collection, DNA extraction, bacterial 16S rRNA sequencing, and taxonomic profiling

[0274]

[0275] Stool samples were collected 1 day before (P0), 1 day after (P1), and 1 month after (P2) carotid artery chemoembolization. Stool samples were collected using a DNA / RNA shielded saliva collection kit and stored at -20°C. Detailed procedures for DNA extraction and 16S rRNA sequencing are described below (DNA extraction, PCR amplification and sequencing, and DNA analysis pipeline). The acquired sequences were analyzed into Operational Taxonomic Units (OTUs), and relative abundance at the species level was calculated as the ratio of the number of OTUs belonging to each species in the phylogenetic tree.

[0276]

[0277] DNA extraction, PCR amplification, and sequencing

[0278]

[0279] A total of 392 stool samples were collected, and 16S rRNA sequencing was performed on 386 samples. Two stool samples were collected at each time point, and samples from three participants were excluded because the OTU readings for samples P0 and P1 were too low to provide sufficient data for analysis. Total DNA was extracted using the FastDNA®SPIN kit for soil (MP BIO, Catalog No.: 6560-200). PCR amplification was performed using primers targeting the V4 region of the 16S rRNA gene. For bacterial amplification, primers 515F (5'-GTGCCAGCMGCCGCGGTAA-3') (Sequence No. 1) and 805R (5'-GGACTACHVGGGTWTCTAAT-3') (Sequence No. 2) were used. The amplification conditions were as follows. Initial denaturation at 95°C for 3 minutes, followed by 25 cycles of denaturation at 95°C for 30 seconds, primer annealing at 55°C for 30 seconds, extension at 72°C for 30 seconds, and finally extension at 72°C for 5 minutes.

[0280] The Nextera XT Index kit V2 (Illumine, Catalog No.: 15052163) was tested as an index primer for library preparation. PCR products were verified using 1% agarose gel electrophoresis and visualized on a GelDoc system. They were purified using HiAccuBead (AccuGene, Catalog No.: ACN01.50). The purified products were pooled at equal concentrations, and short fragments (non-target products) were removed using HiAccuBead. Mixed amplicons were pooled, and sequencing was performed using an Illumina iSeq100 sequencing system (Illumina, USA) at the Asan Institute for Life Sciences (Seoul, Korea). All analyses were performed on an MTP based on EzBioCloud 16S, ChunLab's bioinformatics cloud platform.

[0281]

[0282] DNA analysis pipeline

[0283]

[0284] Raw readout processing using Trimmomatic ver. 0.32 for quality inspection and low quality ( <Q25) 판독 제거로 시작되었다. QC 통과 후, 페어링된 엔드 시퀀스 데이터는 기본 매개변수를 사용하여 VSEARCH 버전 2.13.4의 fastq_mergepairs 명령을 사용하여 병합되었다. 그런 다음 프라이머는 Myers & Miller의 정렬 알고리즘을 사용하여 유사도 컷오프 0.8에서 트리밍했다. 16S rRNA를 인코딩하지 않는 비특이적 앰플리콘은 hmm 프로필을 사용하여 HMMER 소프트웨어 패키지 ver. 3.2.1의 nhmmer에서 감지했다. 고유한 판독이 추출되었고 중복 판독은 VSEARCH의 derep_fulllength 명령을 사용하여 고유한 판독과 클러스터링되었다.

[0285] The EzBioCloud 16S rRNA database was used for classification assignment using the VSEARCH usearch_global command, followed by more accurate pairwise alignment. Chimeric reads with <97% similarity were filtered through reference-based chimeric detection using the UCHIME algorithm and the non-chimeric 16S rRNA database in EzBioCloud. After chimeric filtering, reads (<97% similarity) that were not identified at the species level in the EzBioCloud database were compiled, and de-novo clustering was performed using the cluster_fast command to generate additional OTUs. Finally, OTUs containing a single read (singleton) were excluded from further analysis.

[0286]

[0287] Phylogenetic diversity of the gut microbial community

[0288]

[0289] OTU cluster analysis involved generating taxonomic composition charts for all taxonomic ranks from phylum to species. Alpha-diversity (abundance-based coverage estimators [ACE], Chao, Shannon, and Simpson indices) and beta-diversity (principal coordinate analysis [PCoA]) were performed to measure bacterial diversity within the microbial community.

[0290]

[0291] Reaction evaluation

[0292]

[0293] Tumor response was first evaluated by multiphase CT and / or liver MRI one month after a single session of transarterial chemoembolization. Transarterial chemoembolization was repeated every 6 to 8 weeks while viable hepatocellular carcinoma was present, and follow-up evaluations were performed every 3 months for 2 years when viable hepatocellular carcinoma was absent. After that, evaluations were performed every 6 months. To evaluate treatment response, the modified Response Evaluation Criteria in Solid Tumors (mRECIST) were used (J Hepatol. Feb 2020;72(2):288-306). Responders were defined as patients who achieved complete remission (CR) or partial remission (PR), and non-responders were defined as patients with stable disease (SD) or progressive disease (PD) at the first disease evaluation after transarterial chemoembolization.

[0294]

[0295] Research results

[0296]

[0297] The primary outcome was the association between the initial treatment response to transarterial chemoembolization and changes in microbial diversity and composition. The secondary outcome was the association between the altered gut microbiome and survival outcomes such as overall survival (OS) and progression-free survival (PFS).

[0298]

[0299] Subgroup analysis evaluating the association between dynamic changes in the gut microbial community and survival outcomes

[0300]

[0301] The present invention detected significant differences in the relative abundance of bacterial taxa between responder and non-responder groups by performing a linear discriminant effect size (LEfSe) analysis using an effect size threshold of 3.0 or higher. Patients were assigned to high-abundance and low-abundance subgroups based on the median relative abundance ratio. Then, OS and PFS were compared between the high-abundance and low-abundance subgroups for selected bacterial species, and species with statistically significant differences in survival outcomes between the two subgroups were identified. Dynamic changes in the diversity and relative abundance of these species after transarterial chemoembolization were also analyzed in the responder and non-responder groups.

[0302]

[0303] Statistical analysis

[0304]

[0305] Data were summarized as medians including quartile proportions for continuous variables and as numbers including percentages for categorical variables. Student's t-test or Mann-Whitney U test was used for continuous variables, and chi-square or Fisher's exact test was used for categorical variables. Gut microbial diversity calculation and biomarker discovery programs were performed by ChunLab Inc. in Seoul, Korea, using proprietary software (Journal of Liver Cancer. 3 2020;20(1):32-40). For bacterial diversity analysis, the alpha diversity index was calculated using the R program package 'vegan'. Unweighted and weighted Unifrac distances to assess beta diversity were calculated using the phyloseq package. Taxonomic and functional biomarkers were identified using statistical comparison algorithms (LDA effect size – LEfSe and Kruskal-Wallis H test) with functional profiles predicted by PICRUSt (PLoS Comput Biol. Aug 2009;5(8):e1000465) and the MinPath algorithm (Nat Biotechnol. Sep 2013;31(9):814-21). All of the aforementioned analyses were performed on microbial taxonomic profiling based on ChunLab’s bioinformatics cloud platform, EzBioCloud 16S (Int J Syst Evol Microbiol. May 2017;67(5):1613-1617).

[0306] OS and PFS were estimated using the Kaplan-Meyer method and compared using the log-rank test. Statistical analysis was performed using R software (http: / cran.r-project.org / ). A P-value less than 0.05 was considered statistically significant.

[0307]

[0308] Ethical Statement

[0309]

[0310] Written informed consent was obtained from all participants in this study. The Institutional Review Board of Asan Medical Center (IRB No. 2021-0240) approved the study protocol.

[0311]

[0312] Example 1. Confirmation of reference characteristics and treatment response after carotid artery chemoembolization

[0313]

[0314] The baseline characteristics of the entire cohort are shown in Figures 2a and 2b. The median age was 65.0 [59.0–72.0] years. The majority of patients were male (84.4%) and had chronic hepatitis B (60.4%), which is the cause of hepatocellular carcinoma. At baseline, 51 patients (53.1%) had preserved liver function corresponding to Child-Pugh grade A (53.1%), and 83 patients (86.5%) had cirrhosis. Most patients had very early or early stages of hepatocellular carcinoma (BCLC stage 0 / A [57.3%] versus BCLC stage B [28.1%] and BCLC stage C [14.6%]). The number of previous transarterial chemoembolization sessions prior to the index date ranged from 0 to 9. Prior to initiating transarterial chemoembolization, 27 patients (28.1%) had received prior treatments such as hepatectomy, radiofrequency ablation, radiation therapy, and systemic chemotherapy.

[0315] Initial treatment response after transarterial chemoembolization was classified according to the mRECIST criteria (Figure 3). 63 patients (65.6%) were classified as responders (CR: 26 patients [27.1%]; PR: 37 patients [38.5%]), and 33 patients were non-responders (SD: 27 patients [28.1%]; PD: 6 patients [6.3%]). The objective response rate (ORR) and disease control rate (DCR) at the time of initial response were 65.6% and 93.7%, respectively.

[0316]

[0317] Example 2-1. Confirmation of phylogenetic diversity of gut microbial communities at baseline in initial responders and non-responders

[0318]

[0319] To determine whether the response to transarterial chemoembolization treatment is associated with gut microbiome imbalance, gut metagenome profiles were studied in baseline stool samples (P0) from responders and non-responders. Alpha and beta diversity values ​​were compared to assess the microbial diversity of these two groups. Alpha diversity was analyzed by calculating the abundance-based coverage estimators (ACE), Chao1, Shannon, and Simpson indices (Fig. 4a). The mean alpha diversity was higher in the responder group than in the non-responder group (all P > 0.05). Beta diversity within these two groups was analyzed using the Bray-Curtis distance (Fig. 4b). Significant separation in bacterial community composition was observed between the two groups (p=8e-09). This indicated a distinct difference in microbial communities between the two groups at baseline.

[0320] Therefore, through Example 2-1, it was confirmed that in a prospective study, the gut microbiome of the responder group had greater alpha diversity than that of the non-responder group, and that there was a significant difference in beta diversity between the two groups at baseline. Additionally, alpha diversity in both groups decreased immediately after transarterial chemoembolization and increased after one month, and it was confirmed that the responders showed significant changes in alpha diversity over time, while the changes in beta diversity over time after transarterial chemoembolization were less pronounced compared to the non-responder group.

[0321] In conclusion, early responders to transarterial chemoembolization had higher alpha diversity at baseline and maintained compositional stability within the microbial community throughout treatment compared to non-responders.

[0322]

[0323] Example 2-2. Identification of gut microbial composition and differentially abundant taxa at baseline in initial responders and non-responders.

[0324]

[0325] We evaluated the fecal presence of bacterial taxa that could potentially distinguish the gut microbial communities of the two groups. As shown in Fig. 5a, Firmicutes dominated the microbial communities in both groups, followed by Bacteroidetes, Proteobacteria, Actinobacteriota, and Fusobacteriota. At the family level (Fig. 5b), Prevotellalaceae and Ruminococcaceae were more abundant in the responder group than in the non-responder group, while Lachnospiraceae and Bacteroidaceae were less abundant. At the genus level, Prevotella and Faecalibacterium were more abundant in the responder group than in the non-responder group, while Bacteroides were less abundant (Fig. 5c).

[0326] The proportion of bacterial taxa that dominated the gut microbial community of one group over the other at the species level was evaluated using the phylogenetic tree (Fig. 5d) and the LEfSe method (Fig. 5e). The microbial communities of the responder and non-responder groups were significantly different. Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s were identified as responder-dominant species in the reference stool (P0), whereas Ruminococcus gnavus was a non-responder-dominant species (LEfSe ≥ 3.0, Fig. 5e). The abundance of Roseburia cecicola, Dialister_uc, and DQ797020_s in the responder group was statistically significant (all P < 0.01, Fig. 5f), and the abundance of Ruminococcus gnavus in the non-responder group was similar (p = 0.027, Fig. 5g).

[0327] In conclusion, abundant short-chain fatty acid-producing bacteria were identified at the baseline.

[0328]

[0329] Example 3-1. Confirmation of dynamic changes in gut microbial community diversity over time after transarterial chemoembolization

[0330]

[0331] Changes in microbial community abundance during the post-transarterial chemoembolization period were evaluated in the responder and non-responder groups by comparing alpha diversity (Figs. 6a and 6b). In the responder group, bacterial diversity and abundance decreased significantly the day after transarterial chemoembolization (P1) (P < 0.05 for all alpha diversity indices), but showed a tendency to recover after one month (P2) (P < 0.001 for ACE and CHAO1, Fig. 6a). A similar alpha diversity pattern was observed in the non-responder group, but it was not statistically significant (P > 0.05 for all, Fig. 6b). There were slight changes in beta diversity in both responders and non-responders during the post-transarterial chemoembolization period (Figs. 6c and 6d). However, the patterns differed. The non-responder group showed slightly greater change and variability in microbial composition over time compared to the responder group.

[0332]

[0333] Example 3-2. Identification of candidate indicators of response to transarterial chemoembolization

[0334]

[0335] Roseburia cecicola and PAC001046, identified as dominant tumors in the responder group, significantly decreased after transarterial chemoembolization in the responder group (Fig. 7a). A similar pattern was observed in the non-responder group (although there was no statistically significant change in the case of Roseburia cecicola) (Fig. 7b). Ruminococcus gnavus, recorded as the dominant tumor in the non-responder group, slightly increased after transarterial chemoembolization in the responder group, but tended to decrease over time in the non-responder group (Figs. 7a and 7b). In the responder group, the relative abundance of Dialister_uc was higher than in the non-responder group at baseline, but the abundance consistently decreased over time. After one month, the abundance was significantly lower than P0 and P1 (p=0.019 and 0.043, respectively; Fig. 7a). In contrast, in the non-responder group, the abundance did not change significantly over time (all P > 0.05; Fig. 7b).

[0336] Differences in microbial distribution between the responder and non-responder groups were investigated to determine if there were microorganisms with significantly different abundances between the two groups or potential survival indicators. During a median follow-up period of 6.83 months (95% CI, 5.73–8.43), 16 patients (16.7%) died. The median OS was 23.0 months (95% CI, 13.7–Not estimable, NE) (Fig. 8a). The OS (overall survival) rates at 5, 10, 15, and 25 months were 94.9%, 82.6%, 63.1%, and 39.4%, respectively. Additionally, 46 patients (47.9%) experienced disease progression during the study period, and the median PFS was 7.0 months (95% CI (confidence interval) 6.0–12.1). The incidence rates of PFS at 5, 10, 15, and 25 months were 69.9%, 34.0%, 21.6%, and 11.6%, respectively (Fig. 8b).

[0337]

[0338] The abundance of Roseburia cecicola and Dialister_uc in baseline stools was significantly associated with survival outcomes. Subgroups with an abundance of one of the two species (see Methods) had significantly better OS than subgroups with lower abundance (cut-off values: 0.005 for Roseburia cecicola, 8.0 x 10⁻⁵ for Dialister_uc, Ps < 0.05) (Figs 9a and 9b). Subgroups with low abundance of Roseburia cecicola had a significantly higher risk of death than subgroups with high abundance (hazard ratio [HR] 3.44, 95% CI, 1.10–10.78, p=0.03). The median OS was 13.7 months (95% CI, 10.2-NE, Fig. 9a). For Dialister_uc, the low-abundance subgroup had significantly worse OS than the high-abundance subgroup (HR 3.90, 95% CI, 1.32–11.57, p=0.01). The median OS was 11.1 months (95% CI, 10.2-unestimable, Fig. 9b). For both Dialister_uc and Roseburia cecicola, there was a significant difference in PFS between the low-abundance and high-abundance groups (HR 2.00, 95% CI 1.09–3.67, p=0.03 for Dialister_uc; HR 1.81, 95% CI 1.00–3.31, p=0.05 for Roseburia cecicola, Figs. 9c and 9d).

[0339] As can be seen in Figures 9e and 9f, neither subgroup of Dialister_uc showed significant differences in reference characteristics. In Roseburia cecicola, the high-concentration subgroup had higher BMI and a higher proportion of non-drinkers than the low-concentration subgroup.

[0340] Therefore, through the examples, it was confirmed that the gut microbial community of the responder group at baseline was richer in short-chain fatty acid-producing bacteria than that of the non-responder group. Among these bacterial species, Roseburia cecicola and Dialister_uc were dominant in the responders' baseline stools, and it was confirmed that an abundance of Roseburia cecicola and Dialister_uc at baseline was significantly associated with improved survival outcomes.

[0341] In Ruminococcus gnavus, which was dominant in the reference stools of non-responders, the high concentration group had a longer median OS than the low concentration group (23.0 months [95% CI, 18.8-unestimated] vs. 13.7 months [95% CI, 11.1-NE]). Conversely, the low concentration group had a longer median PFS than the high concentration group (7.2 months [95% CI, 5.47-19.5] vs. 6.9 months [95% CI, 5.67-13.9]). The difference in OS or PFS between the two subgroups was HR 1.42, 95% CI 0.53-3.84, p=0.49 for OS, and HR 0.74, 95% CI 0.41-1.34, p=0.32 for PFS in relation to the abundance subgroup (Figs. 9g and 9h).

[0342]

[0343] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0344] The intestinal microbiota of the present invention serves as a biomarker for predicting the therapeutic responsiveness or prognosis of transarterial chemoembolization in liver cancer patients. The intestinal microbiota of the present invention can be utilized as a potential predictor of clinical outcomes for transarterial chemoembolization, as well as as an adjuvant therapeutic target for the treatment of liver cancer, and can predict the survival rate of liver cancer patients who have undergone transarterial chemoembolization. Therefore, the biomarker of the present invention can be usefully utilized as a marker for predicting the therapeutic responsiveness or prognosis of transarterial chemoembolization in liver cancer patients, and thus has industrial applicability.

Claims

A method for providing information for predicting treatment responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps: (a) Step of separating control group and subject samples; (b) a step of extracting DNA from the above sample and comparing the content of one or more bacteria of a family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroidaceae, one or more bacteria of a genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides, or one or more bacteria of a species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus through metagenome analysis; and (c) If, compared to the control group in step (b) above, the content of one or more bacteria of the family Prevotellaceae and Ruminococcaceae selected from the group consisting of Prevotella and Faecalibacterium, one or more bacteria of the genus selected from the group consisting of Prevotella and Faecalibacterium, or one or more bacteria of the species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s is high, the therapeutic response to transarterial chemoembolization is high, or a good prognosis is predicted, or Step (b) above, in which, compared to the control group, the content of one or more bacteria of the family selected from the group consisting of Lachnospiraceae and Bacteroidaceae, bacteria of the genus Bacteroides, or bacteria of the species Ruminococcus gnavus is high, predicting a low therapeutic response to transarterial chemoembolization or a poor prognosis. In paragraph 1, A method of providing information characterized in that, in step (a) above, the sample is a stool. In paragraph 1, A method for providing information, characterized in that in step (b) above, the metagenomic analysis is performed by performing PCR (polymerase chain reaction) on the extracted DNA using the primer pair of SEQ ID NO. 1 and SEQ ID NO.

2. In paragraph 1, A method for providing information, characterized in that in step (b) above, the metagenomic analysis analyzes the microorganisms of the sample using 16S rRNA sequencing. In paragraph 1, The above information provision method is characterized by analyzing microorganisms of a sample at the species level using LEfSe (linear discriminant analysis (LDA) Effect Size) analysis. In paragraph 5, A method for providing information characterized by predicting high therapeutic responsiveness or a good prognosis based on the linear discriminant analysis (LDA) score of LEfSe analysis, when one or more species of bacteria selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, and DQ797020_s are dominant species among the total microorganisms in a subject sample. In paragraph 5, A method for providing information characterized by predicting that treatment responsiveness is low or the prognosis is poor when Ruminococcus gnavus is the dominant species among the total microorganisms in a subject sample based on the LDA (linear discriminant analysis) score of LEfSe analysis. A method for providing information for predicting treatment responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps: (a) Step of separating control group and subject samples; (b) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from the group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and (c) A step in which, compared to the control group in step (b) above, one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis. A method for providing information for predicting treatment responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps: (S1) Step of separating subject samples before and after carotid artery chemoembolization; (S2) A step of extracting DNA from the above sample and comparing the content of one or more species of bacteria selected from the group consisting of Dialister_uc and Ruminococcus gnavus through metagenome analysis; and (S3) In the above step (S2), if the content of Dialister_uc species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is high or the prognosis is good, or Step (S2) above, if the content of Ruminococcus gnavus species bacteria in the subject sample after transarterial chemoembolization is reduced compared to the subject sample before transarterial chemoembolization, it is predicted that the therapeutic responsiveness to transarterial chemoembolization is low or the prognosis is poor. In Paragraph 9, A method for providing information characterized in that, in the above (S1) step, the sample is a spokesperson. In Paragraph 9, A method for providing information, characterized in that in step (S2) above, the metagenome analysis is performed by performing PCR (polymerase chain reaction) on the extracted DNA using the primer pair of SEQ ID NO. 1 and SEQ ID NO.

2. In Paragraph 9, A method for providing information, characterized in that in step (S2) above, the metagenomic analysis analyzes the microorganisms of the sample using 16S rRNA sequencing. A method for providing information for predicting treatment responsiveness or prognosis for transarterial chemoembolization (TACE), comprising the following steps: (S1) Step of separating control and subject samples after carotid artery chemoembolization; (S2) a step of extracting DNA from the sample and comparing one or more alpha diversity indices selected from a group consisting of ACE (abundance-based coverage estimators) indices, CHAO 1 indices, and Shannon indices through metagenome analysis; and (S3) A step in which, compared to the control group in step (S2), one or more alpha diversity indices selected from the group consisting of a high ACE (abundance-based coverage estimators) index, a high CHAO 1 index, and a high Shannon index are predicted to have a high therapeutic responsiveness to transarterial chemoembolization or a good prognosis. Treatment for transarterial chemoembolization (TACE) with agents measuring the content of one or more bacteria of families selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of genera selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more bacteria of species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus. For reactivity or prognosis prediction purposes. For the preparation of agents for predicting therapeutic response or prognosis for transarterial chemoembolization (TACE), one or more bacteria of the family selected from the group consisting of Prevotellaceae, Ruminococcaceae, Lachnospiraceae, and Bacteroideae; one or more bacteria of the genus selected from the group consisting of Prevotella, Faecalibacterium, and Bacteroides; or one or more species selected from the group consisting of Roseburia cecicola, Dialister_uc, PAC001046, DQ797020_s, and Ruminococcus gnavus Use of a preparation for measuring the content of (species) bacteria.

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