Blood exosomal micro ribonucleic acid profiling reveals the complexity of hepatocellular carcinoma and identifies potential biomarkers for differential diagnosis

BACKGROUND Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths worldwide,but there is a shortage of effective biomarkers for its diagnosis.AIM To explore blood exosomal micro ribonucleic acids(miRNAs)as potential biomarkers for HCC diagnosis.RESULTS The principal component analysis suggested that daily alcohol consumption could alter the blood exosomal miRNA profiles of hepatitis B virus positive non-HCC patients through miR-3168 and miR-223-3p.The miRNA profiles also revealed the tumor stages of HCC patients.High expression of miR-455-5p and miR-30c-5p,which significantly correlated with better overall survival in tumor tissues,could also be detected in blood exosomes.Two pairs of miRNAs(miR-584-5p/miR-106-3p and miR-628-3p/miR-941)showed a 94.1%sensitivity and 68.4%specificity to differentiate HCC patients from non-HCC patients.The specificity of the combination was substantially influenced by alcohol consumption habits.CONCLUSION This study suggested that blood exosomal miRNAs can be used as new noninvasive diagnostic tools for HCC.However,their accuracy could be affected by tumor stage and alcohol consumption habits.

Association between homologous recombination deficiency and outcomes with platinum and platinum-free chemotherapy in patients with triple-negative breast cancer

Objective:The choice of chemotherapeutic regimen for triple-negative breast cancer(TNBC)remains controversial.Homologous recombination deficiency(HRD)has attracted increasing attention in informing chemotherapy treatment.This study was aimed at investigating the feasibility of HRD as a clinically actionable biomarker for platinum-containing and platinum-free therapy.Methods:Chinese patients with TNBC who received chemotherapy between May 1,2008 and March 31,2020 were retrospectively analyzed with a customized 3D-HRD panel.HRD positivity was defined by an HRD score≥30 or deleterious BRCA1/2 mutation.A total of 386 chemotherapy-treated patients with TNBC were screened from a surgical cohort(NCT01150513)and a metastatic cohort,and 189 patients with available clinical and tumor sequencing data were included.Results:In the entire cohort,49.2%(93/189)of patients were identified as HRD positive(40 with deleterious BRCA1/2 mutations and 53 with BRCA1/2 intact with an HRD score of≥30).In the first-line metastatic setting,platinum therapy was associated with longer median progression-free survival(mPFS)than platinum-free therapy[9.1 vs.3.0 months;hazard ratio(HR),0.43;95%confidence interval 0.22–0.84;P=0.01].Among HRD-positive patients,the mPFS was significantly longer in those treated with platinum rather than platinum-free therapy(13.6 vs.2.0 months;HR,0.11;P=0.001).Among patients administered a platinum-free regimen,HRD-negative patients showed a PFS significantly superior to that of HRD-positive patients(P=0.02;treatment-biomarker P-interaction=0.001).Similar results were observed in the BRCA1/2-intact subset.In the adjuvant setting,HRD-positive patients tended to benefit more from platinum chemotherapy than from platinum-free chemotherapy(P=0.05,P-interaction=0.02).Conclusions:HRD characterization may guide decision-making regarding the use of platinum treatment in patients with TNBC in both adjuvant and metastatic settings.

Therapeutic guidance of tumor mutation burden on immune checkpoint inhibitors in advanced non-small cell lung cancer:a systematic review and comprehensive meta-analysis

Background:Tumor mutation burden(TMB)remains a promising but ambiguous predictive biomarker for the efficacy of immune checkpoint inhibitors(ICIs).We investigated the predictive value of TMB in patients with advanced non-small cell lung cancer(NSCLC)treated by ICI-containing therapies under strictly matched clinical settings.Methods:PubMed,Embase,Cochrane Central,ClinicalTrials.gov,and bioRxiv databases were searched till October 16,2021.All randomized controlled trials(RCTs)that compared patients with high TMB(TMB-H)and low TMB(TMB-L)and provided hazard ratio(HR)and corresponding 95%confidence interval(CI)in advanced NSCLC patients receiving ICIs were included,and mirror-based meta-analysis was performed(Part1).Bayesian network meta-analysis was conducted to investigate the efficacy of distinct first-line regimens in TMB-H and TMB-L groups(Part2).Public cohorts were used for validation and further exploration(Part3).Results:Twelve RCTs(n=5527)and 5 public cohorts(n=573)were included.In Part1,TMB-H patients generally exhibited a more significant progression-free survival(PFS)benefit from ICI-containing therapies compared to TMB-L patients(HR=0.58,95%CI:0.49-0.67,P<0.0001).In Part2,anti-PD-1 plus chemotherapy ranked best for PFS in both TMB-H and TMB-L groups.Anti-PD-L1 plus anti-CTLA-4 therapies indicated better PFS and overall survival(OS)benefit than single ICI and chemotherapy in the TMB-H group,but ranked worst in the TMB-L group.Finally,TMB was validated to be an independent predictive biomarker from programmed cell death-ligand 1(PD-L1)expression in Part3,which could further distinguish beneficiaries of ICI-containing therapies with PD-L1<50%.Conclusion:TMB-H could be a predictive biomarker independent of PD-L1 expression to identify beneficiaries of ICI-containing therapy in advanced NSCLC patients.

Bladder preservation in complicated invasive urothelial carcinoma following treatment with cisplatin/gemcitabine plus tislelizumab:A case report

BACKGROUND Invasive urothelial carcinoma(UC)with squamous and glandular differentiation is a highly malignant and complicated pathological subtype,and the standard care is radical cystectomy(RC).However,urinary diversion after RC significantly reduces patient quality of life,thus bladder-sparing therapy has become a research hotspot in this field.Recently,five immune checkpoint inhibitors have been approved for systemic therapy of locally advanced or metastatic bladder cancer by the Food and Drug Administration,but the efficacy of immunotherapy combined with chemotherapy for invasive UC is still unknown,especially for pathological subtypes with squamous and glandular differentiation.CASE SUMMARY We report the case of a 60-year-old male who complained of repetitive painless gross hematuria and was diagnosed with muscle-invasive bladder cancer with squamous and glandular differentiation,defined as cT3N1M0 according to the American Joint Committee on Cancer,who had a strong desire to preserve the bladder.Immunohistochemical staining revealed that programmed cell deathligand 1(PD-L1)expression in the tumor was positive.Thus,a transurethral resection to maximize removal of the bladder tumor was performed under cystoscopy,and the patient subsequently received a combination of chemotherapy(cisplatin/gemcitabine)and immunotherapy(tislelizumab)treatment.No tumor recurrence in the bladder was observed following pathological and imaging examination after 2 cycles and 4 cycles of treatment,respectively.The patient achieved bladder preservation and has been tumor-free for more than two years.CONCLUSION This case shows that the combination of chemotherapy and immunotherapy might be an effective and safe treatment strategy for PD-L1 expression positive UC with divergent histologic differentiation.

Response of human epidermal growth factor receptor 2-positive colorectal cancer to lapatinib monotherapy: A case report

BACKGROUND Human epidermal growth factor receptor 2(HER2)amplification is a molecular driver for a subset of colorectal cancers(CRCs)and one of the major causes of anti-epidermal growth factor receptor(EGFR)treatment failure.Compared to dual anti-HER2 treatments,which have been shown to be effective in HER2-positive metastatic CRC patients,single-agent anti-HER2 therapy is rarely used to treat CRC.CASE SUMMARY Herein,we report a case of RAS/BRAF-wild-type metastatic CRC that was identified as HER2-positive through circulating tumor DNA(ctDNA)testing by next-generation sequencing following the failure of two lines of therapy.Subsequently,the patient was given lapatinib monotherapy that led to a partial response with a progression-free survival of 7.9 mo.Moreover,serial ctDNA detection was used to monitor the efficacy of lapatinib.The aberration of HER2 copy number disappeared when radiographic assessment revealed a partial response.However,a high level of HER2 amplification was detected again at the time of disease progression.Finally,a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha mutation was identified at the time of tumor progression,which may explain the acquired resistance to lapatinib.CONCLUSION This is the first case report of HER2-positive RAS/BRAF wild-type metastatic CRC patient responding to lapatinib monotherapy.It highlights that ctDNA testing is an effective and feasible approach to evaluate the efficacy of anti-HER2 therapy.

Chinese herbal medicine combined with cognitive–behavioural therapy for avoidant paruresis:a controlled trial

Background Avoidant paruresis is a common clinical condition in urology and psychosomatic medicine.However,it has limited treatment options that are safe and effective with few side effects.Aims Our study aimed to investigate the effectiveness and safety of the Chinese herbal Yangxin Tongquan decoction combined with cognitive-behavioural therapy(CBT)for avoidant paruresis.Methods Sixty-eight patients with avoidant paruresis were divided into a treatment group(33 patients)and a control group(35 patients).The control group was assigned 10 weeks of CBT and systematic desensitisation.In addition to CBT and systematic desensitisation,the treatment group was given the Chinese herbal Yangxin Tongquan decoction during the 10-week study.The Shy Bladder Syndrome Scale(SBS)and the Self-rating Anxiety Scale(SAS)were administered before and after treatment to measure any change.Results The overall efficacy in the treatment group(n=30)was 80.0%vs 62.5%in the control group(n=33).Comparing pretreatment and post-treatment measures,both groups showed improvement in SBS scores and SAS scores(treatment group:t_((SBS))=8.397,p_((SBS))<0.001,t_((SAS))=8.216,p_((SAS))<0.001;control group:t_((SBS))=6.802,p_((SBS))<0.001,t_((SAS))=5.171,p_((SAS))<0.001).Moreover,both groups’SBS and SAS scores changed significantly over time(SBS scores:F_(time)=118.299,p<0.001;SAS scores:F_(time)=92.114,p<0.001).However,the treatment group performed better than the control group(SBS scores:F_(time*group)=5.709,p=0.020;SAS scores:F_(time*group)=7.235,p=0.009).Conclusions The Chinese herbal Yangxin Tongquan decoction combined with cognitive-behavioural psychotherapy positively affects the treatment of avoidant paruresis without significant adverse effects.

Relationship of serum polyunsaturated fatty acids with cytokines in colorectal cancer

AIM: To investigate the relationship of serum levels of polyunsaturated fatty acid(PUFA) with kinds of cytokines in colorectal cancer(CRC).METHODS: Serum samples of 100 CRC patients were collected. The concentration of total n-3 PUFA which included C18:3 n-3, C20:5 n-3, C22:5 n-3, C22:6 n-3 and the total n-6 PUFA included C18:2 n-6, C18:3 n-6, C20:3 n-6, C20:4 n-6, and C22:5 n-6 were detected onGC-2010 Plus Gas Chromatograph with a Omegawax TM 250 column. Cytokines were detected by Mag Plex TM-C microspheres. P values for the trend were estimated by creating a continuous variable using the median value within quartiles.RESULTS: Interleukin-6(IL-6) showed significantly positive association with the C20:4 n-6(P for trend = 0.004). Interferon gamma(IFN-γ) showed significant positive association with the C22:5 n-3(P for trend = 0.035). IL-8 and matrix metalloproteinase-9(MMP-9) showed significant inverse association with the C22:6 n-3(P for trend = 0.049, and 0.021). MMP-2 showed significant inverse association with the C20:5 n-3(P for trend = 0.008). MMP-7 showed significantly positive association with the ratio of n-6 PUFA and n-3 PUFA(P for trend = 0.008). MMP-7 also showed significantly inverse association with the ratio of C20:4 n-6 and(n-6 PUFA + n-3 PUFA)(P for trend = 0.024). IL-10(P for trend = 0.023) and IL-6(P for trend = 0.036) showed significantly positive association with the ratio of C20:4 n-6 and C20:5 n-3.CONCLUSION: Our data suggested that serum levels of PUFA is related to the inflammation of CRC, and also play different role in regulation of immune response.

Treatment with sorafenib plus camrelizumab after splenectomy for primary splenic angiosarcoma with liver metastasis:A case report and literature review

BACKGROUND Primary splenic angiosarcoma(PSA)is an extremely rare and aggressive mesenchymal malignancy with high metastatic potential and a poor prognosis.There are no established treatment guidelines for PSA,even for adjuvant therapy.This rare case may provide a reliable therapeutic regime for a better prognosis.CASE SUMMARY A 49-year-old female who complained of right-upper quadrant abdominal pain was diagnosed as having PSA with splenic rupture and liver metastasis.After splenectomy and liver tumor resection,she received sorafenib and camrelizumab therapy.After 15 mo of follow-up,she is in good condition,without recurrence or any identified metastasis.CONCLUSION Immunotherapy combined with targeted therapy could be a potential option for the adjuvant therapy of PSA.

The tumor immune microenvironment transcriptomic subtypes of colorectal cancer for prognosis and development of precise immunotherapy

Background:Biomarkers based on immune context may guide prognosis prediction.T-cell inactivation,exclusion,or dysfunction could cause unfavorable tumor microenvironments,which affect immunotherapy and prognosis.However,none of the immuno-biomarkers reported to date can differentiate colorectal-cancer(CRC)patients.Thus,we aimed to classify CRC patients according to the levels of T-cell activation,exclusion,and dysfunction in the tumor microenvironment.Methods:RNAseq data of 618 CRC patients from The Cancer Genome Atlas and microarray data of 316 CRC patients from Gene Expression Omnibus were analysed using the Tumor Immune Dysfunction and Exclusion algorithm.Unsupervised clustering was used to classify patients.Results:Based on the expression signatures of myeloid-derived suppressor cells,cancer-associated fibroblasts,M2-like tumor-associated macrophages,cytotoxic T-lymphocytes,and PD-L1,all patients were clustered into four subtypes:cluster 1 had a high level of immune dysfunction,cluster 2 had a low level of immune activation,cluster 3 had intense immune exclusion,and cluster 4 had a high level of immune activation and a moderate level of both dysfunction and exclusion signatures.Compared with cluster 1,the hazard ratios and 95%confidential intervals for overall survival were 0.63(0.35-1.13)for cluster 2,0.55(0.29-1.03)for cluster 3,and 0.30(0.14-0.64)for cluster 4 in multivariate Cox regression.Similar immune clustering and prognosis patterns were obtained upon validation in the GSE39582 cohort.In subgroup analysis,immune clustering was significantly associated with overall survival among stage I/II patients,microsatellite stable/instability-low patients,and patients not treated with adjuvant therapy.Conclusions:Our findings demonstrated that classifying CRC patients into different immune subtypes serves as a reliable prognosis predictor and may help to refine patient selection for personalized cancer immunotherapy.

A case report of response to crizotinib in chemotherapy-refractory metastatic gallbladder cancer with met amplification and acquired resistance resulting from the loss of MET amplification

Gallbladder cancer(GBC)is a highly invasive disease and the most prevalent malignancy of the biliary system.Patients with GBC are commonly diagnosed at a late stage and have an unfavorable prognosis.Palliative chemotherapy has been the standard care for recurrent or metastatic disease in the past decades.Recently,several targeted therapies have been investigated in advanced biliary tract cancer(BTC)including inhibitors of genes or pathways such as FGFR2 fusions or rearrangements,IDH1 mutations,and NTRK gene fusions.Also,several clinical studies involving molecular stratification have been performed in defined patient groups,for example,BRAF V600E and HER2.Mesenchymal epithelial transition(MET)encodes a tyrosine kinase receptor and its ligand hepatocyte growth factor is a proto-oncogene.Targeting the MET signaling pathway is an effective strategy in numerous cancer types.However,the poor efficacy of MET inhibitors has been demonstrated in several phase II studies,but currently no reports have explained the potential mechanisms of resistance to MET inhibitors in BTC.In this article,we report a case of metastatic GBC with MET amplification that exhibited a rapid response to crizotinib after the failure of two lines of chemotherapy.After the patient had progressed and discontinued crizotinib,cabozantinib was introduced.Analysis of circulating tumor DNA(ctDNA)by nextgeneration sequencing(NGS)indicated a loss of MET amplification status.To our knowledge,this is the first case study demonstrating the use of NGS in ctDNA to monitor the development of acquired resistance during anti-MET treatment in GBC.

mir-3168 targeted inhibition of TP53 promotes malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells

Objective:To investigate the effect of mir-3168 on the malignant transformation and cisplatin resistance of AGS and AGS/DDP gastric cancer cells,and to verify its target gene.Methods:The expression of mir-3168 in AGS and AGS/DDP gastric cancer cells was detected by qPCR,and mir-3168 mimic,inhibitor and negative control were synthesized.They were transfected into AGS and AGS/DDP gastric cancer cells,respectively.The expression of mir-3168 and TP53 mRNA was detected by qPCR.Cell viability was detected by CCK8 under gradient cisplatin treatment and non treatment,apoptosis was detected by flow cytometry,cell invasion was detected by Transwell,and TP53 protein expression was detected by western blot,The database predicted the binding sites of mir-3168 and TP53.According to the binding sites,the double luciferase experiment was used to verify the binding of mir-3168 and TP53.Results:Compared with cisplatin sensitive gastric cancer cell AGS,mir-3168 was significantly overexpressed in cisplatin resistant gastric cancer cell AGS/DDP;mir-3168 mimic promotes cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and inhibits apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 inhibitor inhibits cisplatin resistance,proliferation and invasion of AGS and AGS/DDP gastric cancer cells,and promotes apoptosis of AGS and AGS/DDP gastric cancer cells;mir-3168 mimic inhibits the expression of TP53 mRNA and protein,and mir-3168 inhibitor promotes the expression of TP53 mRNA and protein;Targetscan database predicted that there was a binding point between mir-3168 and TP53,and the double luciferase experiment suggested that mir-3168 was bound to TP53 through the predicted binding site.Conclusion:mir-3168 may promote the malignant transformation of AGS and AGS/DDP gastric cancer cells and cisplatin resistance by targeting TP53.

Characterization of DNA damage response deficiency in pancreatic cancer patients from China

To the editor,Pancreatic ductal adenocarcinoma(PDAC)has the worst prognosis among all common malignant solid tumors,with a 5-year overall survival(OS)rate of less than 10%[1].Few effective targets for anticancer ther-apy have been confirmed in pancreatic cancer.Recently,it was substantiated that pancreatic cancer patients carry-ing deleterious mutations of the DNA damage response(DDR)genes are more likely to benefit from platinum-based chemotherapy[2]and poly(adenosine diphosphate-ribose)polymerase(PARP)inhibitor[3].