HepG2 exosomes coated luteolin nanoparticles remodeling hepatic stellate cells and combination with sorafenib for the treatment of hepatocellular carcinoma

Abou-Alfa GK, Johnson P, Knox JJ, Capanu M, Davidenko I, Lacava J, Leung T, Gansukh B, Saltz LB (2010) Doxorubicin plus sorafenib vs doxorubicin alone in patients with advanced hepatocellular carcinoma: a randomized trial. JAMA 304:2154–2160. https://doi.org/10.1001/jama.2010.1672 Agrawal AK, Aqil F, Jeyabalan J, Spencer WA, Beck J, Gachuki BW, Alhakeem SS, Oben K, Munagala R, Bondada S, Gupta RC (2017) Milk-derived exosomes for oral delivery of paclitaxel. Nanomedicine 13:1627–1636. https://doi.org/10.1016/j.nano.2017.03.001 Ashour AA, El-Kamel AH, Mehanna RA, Mourad G, Heikal LA (2022) Luteolin-loaded exosomes derived from bone marrow mesenchymal stem cells: a promising therapy for liver fibrosis. Drug Deliv 29:3270–3280. https://doi.org/10.1080/10717544.2022.2142700 Azzariti A, Mancarella S, Porcelli L, Quatrale AE, Caligiuri A, Lupo L, Dituri F, Giannelli G (2016) Hepatic stellate cells induce hepatocellular carcinoma cell resistance to sorafenib through the laminin-332/α3 integrin axis recovery of focal adhesion kinase ubiquitination. Hepatology 64:2103–2117. https://doi.org/10.1002/hep.28835 Bárcena C, Stefanovic M, Tutusaus A, Martinez-Nieto GA, Martinez L, García-Ruiz C, de Mingo A, Caballeria J, Fernandez-Checa JC, Marí M, Morales A (2015) Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer. Sci Rep 5:7916. https://doi.org/10.1038/srep07916 Chakrabarti M, Ray SK (2015) Synergistic anti-tumor actions of luteolin and silibinin prevented cell migration and invasion and induced apoptosis in glioblastoma SNB19 cells and glioblastoma stem cells. Brain Res 1629:85–93. https://doi.org/10.1016/j.brainres.2015.10.010 Chang L-W, Hou M-L, Tsai T-H (2014) Silymarin in liposomes and ethosomes: pharmacokinetics and tissue distribution in free-moving rats by high-performance liquid chromatography-tandem mass spectrometry. J Agric Food Chem 62:11657–11665. https://doi.org/10.1021/jf504139g Chen Q, Chen J, Yang Z, Xu J, Xu L, Liang C, Han X, Liu Z (2019) Nanoparticle-enhanced radiotherapy to trigger robust cancer immunotherapy. Adv Mater 31:e1802228. https://doi.org/10.1002/adma.201802228 Chen L, Chang S, Zhao L, Li B, Zhang S, Yun C, Wu X, Meng J, Li G, Guo S, Duan J (2022) Biosynthesis of a water solubility-enhanced succinyl glucoside derivative of luteolin and its neuroprotective effect. Microb Biotechnol 15:2401–2410. https://doi.org/10.1111/1751-7915.14095 Copp JA, Fang RH, Luk BT, Hu C-MJ, Gao W, Zhang K, Zhang L (2014) Clearance of pathological antibodies using biomimetic nanoparticles. Proc Natl Acad Sci USA 111:13481–13486. https://doi.org/10.1073/pnas.1412420111 Cummins CB, Wang X, Nunez Lopez O, Graham G, Tie H-Y, Zhou J, Radhakrishnan RS (2018) Luteolin-mediated inhibition of hepatic stellate cell activation via suppression of the STAT3 pathway. Int J Mol Sci 19:1567. https://doi.org/10.3390/ijms19061567 D’Ambrosio DN, Walewski JL, Clugston RD, Berk PD, Rippe RA, Blaner WS (2011) Distinct populations of hepatic stellate cells in the mouse liver have different capacities for retinoid and lipid storage. PLoS ONE 6:e24993. https://doi.org/10.1371/journal.pone.0024993 Eisai and merck and CO., INC (2018) kenilworth, N.J., U.S.A. announce FDA approval of lenvima® (lenvatinib) capsules for first-line treatment of unresectable hepatocellular carcinoma (HCC), news release: 2018, Eisai Co., Ltd. https://www.eisai.com/news/2018/news201870.html. Accessed 2 July 2023 European Association For The Study Of The Liver, European Organisation For Research And Treatment Of Cancer (2012) EASL-EORTC clinical practice guidelines: management of hepatocellular carcinoma. J Hepatol 56:908–943. https://doi.org/10.1016/j.jhep.2011.12.001 Fang RH, Hu C-MJ, Luk BT, Gao W, Copp JA, Tai Y, O’Connor DE, Zhang L (2014) Cancer cell membrane-coated nanoparticles for anticancer vaccination and drug delivery. Nano Lett 14:2181–2188. https://doi.org/10.1021/nl500618u Faruqu FN, Xu L, Al-Jamal KT (2018) Preparation of exosomes for siRNA delivery to cancer cells. J vis Exp. https://doi.org/10.3791/58814 Feng X-Q, Rong L-W, Wang R-X, Zheng X-L, Zhang L, Zhang L, Lin Y, Wang X, Li Z-P (2018) Luteolin and sorafenib combination kills human hepatocellular carcinoma cells through apoptosis potentiation and JNK activation. Oncol Lett 16:648–653. https://doi.org/10.3892/ol.2018.8640 Feng T, Fang F, Zhang C, Li T, He J, Shen Y, Yu H, Liu X (2022) Fluid shear stress-induced exosomes from liver cancer cells promote activation of cancer-associated fibroblasts via IGF2-PI3K axis. Front Biosci (landmark Ed). 27:104. https://doi.org/10.31083/j.fbl2703104 Friedman SL (2008) Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver. Physiol Rev 88:125–172. https://doi.org/10.1152/physrev.00013.2007 Grandhi MS, Kim AK, Ronnekleiv-Kelly SM, Kamel IR, Ghasebeh MA, Pawlik TM (2016) Hepatocellular carcinoma: from diagnosis to treatment. Surg Oncol 25:74–85. https://doi.org/10.1016/j.suronc.2016.03.002 Han Z, Lv W, Li Y, Chang J, Zhang W, Liu C, Sun J (2020) Improving tumor targeting of exosomal membrane-coated polymeric nanoparticles by conjugation with aptamers. ACS Appl Bio Mater 3:2666–2673. https://doi.org/10.1021/acsabm.0c00181 Haney MJ, Klyachko NL, Zhao Y, Gupta R, Plotnikova EG, He Z, Patel T, Piroyan A, Sokolsky M, Kabanov AV, Batrakova EV (2015) Exosomes as drug delivery vehicles for Parkinson’s disease therapy. J Control Release 207:18–30. https://doi.org/10.1016/j.jconrel.2015.03.033 Higashi T, Friedman SL, Hoshida Y (2017) Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev 121:27–42. https://doi.org/10.1016/j.addr.2017.05.007 Hu C-MJ, Zhang L, Aryal S, Cheung C, Fang RH, Zhang L (2011) Erythrocyte membrane-camouflaged polymeric nanoparticles as a biomimetic delivery platform. Proc Natl Acad Sci USA 108:10980–10985. https://doi.org/10.1073/pnas.1106634108 Khawar IA, Park JK, Jung ES, Lee MA, Chang S, Kuh H-J (2018) Three dimensional mixed-cell spheroids mimic stroma-mediated chemoresistance and invasive migration in hepatocellular carcinoma. Neoplasia 20:800–812. https://doi.org/10.1016/j.neo.2018.05.008 Kim MS, Haney MJ, Zhao Y, Mahajan V, Deygen I, Klyachko NL, Inskoe E, Piroyan A, Sokolsky M, Okolie O, Hingtgen SD, Kabanov AV, Batrakova EV (2016) Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells. Nanomedicine 12:655–664. https://doi.org/10.1016/j.nano.2015.10.012 Kouwaki T, Fukushima Y, Daito T, Sanada T, Yamamoto N, Mifsud EJ, Leong CR, Tsukiyama-Kohara K, Kohara M, Matsumoto M, Seya T, Oshiumi H (2016) Extracellular vesicles including exosomes regulate innate immune responses to hepatitis B virus infection. Front Immunol 7:335. https://doi.org/10.3389/fimmu.2016.00335 Lee Y, Kwon YH (2019) Regulation of apoptosis and autophagy by luteolin in human hepatocellular cancer Hep3B cells. Biochem Biophys Res Commun 517:617–622. https://doi.org/10.1016/j.bbrc.2019.07.073 Lee NP, Chan CM, Tung LN, Wang HK, Law S (2018) Tumor xenograft animal models for esophageal squamous cell carcinoma. J Biomed Sci 25:66. https://doi.org/10.1186/s12929-018-0468-7 Lenggenhager D, Amrutkar M, Sántha P, Aasrum M, Löhr J-M, Gladhaug IP, Verbeke CS (2019) Commonly used pancreatic stellate cell cultures differ phenotypically and in their interactions with pancreatic cancer cells. Cells 8:23. https://doi.org/10.3390/cells8010023 Li T, Shi Z, Rockey DC (2012) Preproendothelin-1 expression is negatively regulated by IFNγ during hepatic stellate cell activation. Am J Physiol Gastrointest Liver Physiol 302:G948-957. https://doi.org/10.1152/ajpgi.00359.2011 Li L, Piontek K, Ishida M, Fausther M, Dranoff JA, Fu R, Mezey E, Gould SJ, Fordjour FK, Meltzer SJ, Sirica AE, Selaru FM (2017) Extracellular vesicles carry microRNA-195 to intrahepatic cholangiocarcinoma and improve survival in a rat model. Hepatology 65:501–514. https://doi.org/10.1002/hep.28735 Li S, Wu Y, Ding F, Yang J, Li J, Gao X, Zhang C, Feng J (2020) Engineering macrophage-derived exosomes for targeted chemotherapy of triple-negative breast cancer. Nanoscale 12:10854–10862. https://doi.org/10.1039/d0nr00523a Li H, Wang X, Guo X, Wan Q, Teng Y, Liu J (2022) Development of rapamycin-encapsulated exosome-mimetic nanoparticles-in-PLGA microspheres for treatment of hemangiomas. Biomed Pharmacother 148:112737. https://doi.org/10.1016/j.biopha.2022.112737 Li Y, Wu J, Qiu X, Dong S, He J, Liu J, Xu W, Huang S, Hu X, Xiang D-X (2023) Bacterial outer membrane vesicles-based therapeutic platform eradicates triple-negative breast tumor by combinational photodynamic/chemo-/immunotherapy. Bioact Mater 20:548–560. https://doi.org/10.1016/j.bioactmat.2022.05.037 Ligorio M, Sil S, Malagon-Lopez J, Nieman LT, Misale S, Di Pilato M, Ebright RY, Karabacak MN, Kulkarni AS, Liu A, Vincent Jordan N, Franses JW, Philipp J, Kreuzer J, Desai N, Arora KS, Rajurkar M, Horwitz E, Neyaz A, Tai E, Magnus NKC, Vo KD, Yashaswini CN, Marangoni F, Boukhali M, Fatherree JP, Damon LJ, Xega K, Desai R, Choz M, Bersani F, Langenbucher A, Thapar V, Morris R, Wellner UF, Schilling O, Lawrence MS, Liss AS, Rivera MN, Deshpande V, Benes CH, Maheswaran S, Haber DA, Fernandez-Del-Castillo C, Ferrone CR, Haas W, Aryee MJ, Ting DT (2019) Stromal microenvironment shapes the intratumoral architecture of pancreatic cancer. Cell 178:160–175. https://doi.org/10.1016/j.cell.2019.05.012 Lin H, Zhang R, Wu W, Lei L (2021) miR-4454 promotes hepatic carcinoma progression by targeting Vps4A and Rab27A. Oxid Med Cell Longev 2021:9230435. https://doi.org/10.1155/2021/9230435 Liu L, Cao Y, Chen C, Zhang X, McNabola A, Wilkie D, Wilhelm S, Lynch M, Carter C (2006) Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res 66:11851–11858. https://doi.org/10.1158/0008-5472.CAN-06-1377 Liu Y, Yue H, Xu S, Wang F, Ma N, Li K, Qiao L, Wang J (2015) First-line gemcitabine and oxaliplatin (GEMOX) plus sorafenib, followed by sorafenib as maintenance therapy, for patients with advanced hepatocellular carcinoma: a preliminary study. Int J Clin Oncol 20:952–959. https://doi.org/10.1007/s10147-015-0796-5 Liu C, Zhang W, Li Y, Chang J, Tian F, Zhao F, Ma Y, Sun J (2019a) Microfluidic sonication to assemble exosome membrane-coated nanoparticles for immune evasion-mediated targeting. Nano Lett 19:7836–7844. https://doi.org/10.1021/acs.nanolett.9b02841 Liu J, Chang B, Li Q, Xu L, Liu X, Wang G, Wang Z, Wang L (2019b) Redox-responsive dual drug delivery nanosystem suppresses cancer repopulation by abrogating doxorubicin-promoted cancer stemness, metastasis, and drug resistance. Adv Sci (weinh) 6:1801987. https://doi.org/10.1002/advs.201801987 Liu Y, Wu X, Chen F, Li H, Wang T, Liu N, Sun K, Zhou G, Tao K (2022) Modulating cancer-stroma crosstalk by a nanoparticle-based photodynamic method to pave the way for subsequent therapies. Biomaterials 289:121813. https://doi.org/10.1016/j.biomaterials.2022.121813 Luo N, Li J, Chen Y, Xu Y, Wei Y, Lu J, Dong R (2021) Hepatic stellate cell reprogramming via exosome-mediated CRISPR/dCas9-VP64 delivery. Drug Deliv 28:10–18. https://doi.org/10.1080/10717544.2020.1850917 Ma X, Qiu Y, Sun Y, Zhu L, Zhao Y, Li T, Lin Y, Ma D, Qin Z, Sun C, Han L (2020) NOD2 inhibits tumorigenesis and increases chemosensitivity of hepatocellular carcinoma by targeting AMPK pathway. Cell Death Dis 11:174. https://doi.org/10.1038/s41419-020-2368-5 Maheswaran T, Rushbrook SM (2012) Epithelial–mesenchymal transition and the liver: role in hepatocellular carcinoma and liver fibrosis. J Gastroenterol Hepatol 27:418–420. https://doi.org/10.1111/j.1440-1746.2012.07060.x Marcucci F, Stassi G, De Maria R (2016) Epithelial–mesenchymal transition: a new target in anticancer drug discovery. Nat Rev Drug Discov 15:311–325. https://doi.org/10.1038/nrd.2015.13 Mathieu M, Névo N, Jouve M, Valenzuela JI, Maurin M, Verweij FJ, Palmulli R, Lankar D, Dingli F, Loew D, Rubinstein E (2021) Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9. Nat Commun. https://doi.org/10.1038/s41467-021-24384-2 Niu G, Zhang X, Hong R, Yang X, Gu J, Song T, Hu Z, Chen L, Wang X, Xia J, Ke Z, Ren J, Hong L (2021) GJA1 promotes hepatocellular carcinoma progression by mediating TGF-β-induced activation and the epithelial-mesenchymal transition of hepatic stellate cells. Open Med (wars) 16:1459–1471. https://doi.org/10.1515/med-2021-0344 Onzi GR, Ledur PF, Hainzenreder LD, Bertoni APS, Silva AO, Lenz G, Wink MR (2016) Analysis of the safety of mesenchymal stromal cells secretome for glioblastoma treatment. Cytotherapy 18:828–837. https://doi.org/10.1016/j.jcyt.2016.03.299 Pei W, Li X, Bi R, Zhang X, Zhong M, Yang H, Zhang Y, Lv K (2021) Exosome membrane-modified M2 macrophages targeted nanomedicine: treatment for allergic asthma. J Control Release 338:253–267. https://doi.org/10.1016/j.jconrel.2021.08.024 Pinto M, Silva V, Barreiro S, Silva R, Remião F, Borges F, Fernandes C (2022) Brain drug delivery and neurodegenerative diseases: polymeric PLGA-based nanoparticles as a forefront platform. Ageing Res Rev 79:101658. https://doi.org/10.1016/j.arr.2022.101658 Pittala S, Krelin Y, Shoshan-Barmatz V (2018) Targeting liver cancer and associated pathologies in mice with a mitochondrial VDAC1-based peptide. Neoplasia 20:594–609. https://doi.org/10.1016/j.neo.2018.02.012 Prieto-Domínguez N, Ordóñez R, Fernández A, Méndez-Blanco C, Baulies A, Garcia-Ruiz C, Fernández-Checa JC, Mauriz JL, González-Gallego J (2016) Melatonin-induced increase in sensitivity of human hepatocellular carcinoma cells to sorafenib is associated with reactive oxygen species production and mitophagy. J Pineal Res 61:396–407. https://doi.org/10.1111/jpi.12358 Qiu Z, Li H, Zhang Z, Zhu Z, He S, Wang X, Wang P, Qin J, Zhuang L, Wang W, Xie F, Gu Y, Zou K, Li C, Li C, Wang C, Cen J, Chen X, Shu Y, Zhang Z, Sun L, Min L, Fu Y, Huang X, Lv H, Zhou H, Ji Y, Zhang Z, Meng Z, Shi X, Zhang H, Li Y, Hui L (2019) A pharmacogenomic landscape in human liver cancers. Cancer Cell 36:179-193.e11. https://doi.org/10.1016/j.ccell.2019.07.001 Rangwala F, Williams KP, Smith GR, Thomas Z, Allensworth JL, Lyerly HK, Diehl AM, Morse MA, Devi GR (2012) Differential effects of arsenic trioxide on chemosensitization in human hepatic tumor and stellate cell lines. BMC Cancer 12:402. https://doi.org/10.1186/1471-2407-12-402 Ruan C, Liu L, Lu Y, Zhang Y, He X, Chen X, Zhang Y, Chen Q, Guo Q, Sun T, Jiang C (2018) Substance P-modified human serum albumin nanoparticles loaded with paclitaxel for targeted therapy of glioma. Acta Pharm Sin B 8:85–96. https://doi.org/10.1016/j.apsb.2017.09.008 Ruan Q, Wang H, Burke LJ, Bridle KR, Li X, Zhao C-X, Crawford DHG, Roberts MS, Liang X (2020) Therapeutic modulators of hepatic stellate cells for hepatocellular carcinoma. Int J Cancer 147:1519–1527. https://doi.org/10.1002/ijc.32899 Ruman U, Fakurazi S, Masarudin MJ, Hussein MZ (2020) Nanocarrier-based therapeutics and theranostics drug delivery systems for next generation of liver cancer nanodrug modalities. Int J Nanomedicine 15:1437–1456. https://doi.org/10.2147/IJN.S236927 Seitz T, Freese K, Dietrich P, Thasler WE, Bosserhoff A, Hellerbrand C (2020) Fibroblast growth factor 9 is expressed by activated hepatic stellate cells and promotes progression of hepatocellular carcinoma. Sci Rep 10:4546. https://doi.org/10.1038/s41598-020-61510-4 Shany S, Bernheimer AW, Grushoff PS, Kim KS (1974) Evidence for membrane cholesterol as the common binding site for cereolysin, streptolysin O and saponin. Mol Cell Biochem 3:179–186. https://doi.org/10.1007/BF01686643 Shiraha H, Iwamuro M, Okada H (2020) Hepatic stellate cells in liver tumor. Adv Exp Med Biol 1234:43–56. https://doi.org/10.1007/978-3-030-37184-5_4 Sieghart W, Pinter M, Dauser B, Rohr-Udilova N, Piguet A-C, Prager G, Hayden H, Dienes H-P, Dufour J-F, Peck-Radosavljevic M (2012) Erlotinib and sorafenib in an orthotopic rat model of hepatocellular carcinoma. J Hepatol 57:592–599. https://doi.org/10.1016/j.jhep.2012.04.034 Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71:209–249. https://doi.org/10.3322/caac.21660 Tang W, Chen Z, Zhang W, Cheng Y, Zhang B, Wu F, Wang Q, Wang S, Rong D, Reiter FP, De Toni EN, Wang X (2020) The mechanisms of sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects. Signal Transduct Target Ther 5:87. https://doi.org/10.1038/s41392-020-0187-x Théry C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol. https://doi.org/10.1002/0471143030.cb0322s30 Verma VK, Li H, Wang R, Hirsova P, Mushref M, Liu Y, Cao S, Contreras PC, Malhi H, Kamath PS, Gores GJ, Shah VH (2016) Alcohol stimulates macrophage activation through caspase-dependent hepatocyte derived release of CD40L containing extracellular vesicles. J Hepatol 64:651–660. https://doi.org/10.1016/j.jhep.2015.11.020 Wang Z, Zhao Z, Wu T, Song L, Zhang Y (2015a) Sorafenib-irinotecan sequential therapy augmented the anti-tumor efficacy of monotherapy in hepatocellular carcinoma cells HepG2. Neoplasma 62:172–179. https://doi.org/10.4149/neo_2015_022 Wang Y, Li H, Liang Q, Liu B, Mei X, Ma Y (2015b) Combinatorial immunotherapy of sorafenib and blockade of programmed death-ligand 1 induces effective natural killer cell responses against hepatocellular carcinoma. Tumour Biol 36:1561–1566. https://doi.org/10.1007/s13277-014-2722-2 Wang F, Li L, Piontek K, Sakaguchi M, Selaru FM (2018a) Exosome miR-335 as a novel therapeutic strategy in hepatocellular carcinoma. Hepatology 67:940–954. https://doi.org/10.1002/hep.29586 Wang S, Xu M, Li X, Su X, Xiao X, Keating A, Zhao RC (2018b) Exosomes released by hepatocarcinoma cells endow adipocytes with tumor-promoting properties. J Hematol Oncol 11:82. https://doi.org/10.1186/s13045-018-0625-1 Wang X, Xiang Z, Liu Y, Huang C, Pei Y, Wang X, Zhi H, Wong WH-S, Wei H, Ng IO-L, Lee PP-W, Chan GC-F, Lau Y-L, Tu W (2020) Exosomes derived from Vδ2-T cells control Epstein-Barr virus-associated tumors and induce T cell antitumor immunity. Sci Transl Med. 12:eaaz3426. https://doi.org/10.1126/scitranslmed.aaz3426 Wang Y-T, Cai M-D, Sun L-L, Hua R-N (2021a) A rapid and facile separation-detection integrated strategy for exosome profiling based on boronic acid-directed coupling immunoaffinity. Anal Chem 93:16059–16067. https://doi.org/10.1021/acs.analchem.1c03643 Wang J, Chen D, Ho EA (2021b) Challenges in the development and establishment of exosome-based drug delivery systems. J Control Release 329:894–906. https://doi.org/10.1016/j.jconrel.2020.10.020 Wang X, Zhang Y, Mu X, Tu CR, Chung Y, Tsao SW, Chan GC-F, Leung W-H, Lau Y-L, Liu Y, Tu W (2022) Exosomes derived from γδ-T cells synergize with radiotherapy and preserve antitumor activities against nasopharyngeal carcinoma in immunosuppressive microenvironment. J Immunother Cancer 10:e003832. https://doi.org/10.1136/jitc-2021-003832 Wilhelm SM, Carter C, Tang L, Wilkie D, McNabola A, Rong H, Chen C, Zhang X, Vincent P, McHugh M, Cao Y, Shujath J, Gawlak S, Eveleigh D, Rowley B, Liu L, Adnane L, Lynch M, Auclair D, Taylor I, Gedrich R, Voznesensky A, Riedl B, Post LE, Bollag G, Trail PA (2004) BAY 43–9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis. Cancer Res 64:7099–7109. https://doi.org/10.1158/0008-5472.CAN-04-1443 Witwer KW, Buzás EI, Bemis LT, Bora A, Lässer C, Lötvall J, Nolte-’t Hoen EN, Piper MG, Sivaraman S, Skog J, Théry C, Wauben MH, Hochberg F (2013) Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J Extracell Vesicles. https://doi.org/10.3402/jev.v2i0.20360 Wu J-Y, Li Y-J, Hu X-B, Huang S, Xiang D-X (2021) Preservation of small extracellular vesicles for functional analysis and therapeutic applications: a comparative evaluation of storage conditions. Drug Deliv 28:162–170. https://doi.org/10.1080/10717544.2020.1869866 Xia S, Pan Y, Liang Y, Xu J, Cai X (2020) The microenvironmental and metabolic aspects of sorafenib resistance in hepatocellular carcinoma. EBioMedicine 51:102610. https://doi.org/10.1016/j.ebiom.2019.102610 Xia C, Dong X, Li H, Cao M, Sun D, He S, Yang F, Yan X, Zhang S, Li N, Chen W (2022) Cancer statistics in China and United States, 2022: profiles, trends, and determinants. Chin Med J (engl) 135:584–590. https://doi.org/10.1097/CM9.0000000000002108 Xiao W, Dong W, Zhang C, Saren G, Geng P, Zhao H, Li Q, Zhu J, Li G, Zhang S, Ye M (2013) Effects of the epigenetic drug MS-275 on the release and function of exosome-related immune molecules in hepatocellular carcinoma cells. Eur J Med Res 18:61. https://doi.org/10.1186/2047-783X-18-61 Xie Z, Gao Y, Ho C, Li L, Jin C, Wang X, Zou C, Mao Y, Wang X, Li Q, Fu D, Zhang Y-F (2022) Exosome-delivered CD44v6/C1QBP complex drives pancreatic cancer liver metastasis by promoting fibrotic liver microenvironment. Gut 71:568–579. https://doi.org/10.1136/gutjnl-2020-323014 Xin H, Li Y, Chopp M (2014) Exosomes/miRNAs as mediating cell-based therapy of stroke. Front Cell Neurosci 8:377. https://doi.org/10.3389/fncel.2014.00377 Xu J, Wan Z, Tang M, Lin Z, Jiang S, Ji L, Gorshkov K, Mao Q, Xia S, Cen D, Zheng J, Liang X, Cai X (2020) N6-methyladenosine-modified CircRNA-SORE sustains sorafenib resistance in hepatocellular carcinoma by regulating β-catenin signaling. Mol Cancer 19:163. https://doi.org/10.1186/s12943-020-01281-8 Xu H, Liao C, Liang S, Ye B-C (2021) A novel peptide-equipped exosomes platform for delivery of antisense oligonucleotides. ACS Appl Mater Interfaces 13:10760–10767. https://doi.org/10.1021/acsami.1c00016 Xue F, Liu Y, Chu H, Wen Y, Yan L, Tang Q, Xiao E, Zhang D, Zhang H (2016) eIF5A2 is an alternative pathway for cell proliferation in cetuximab-treated epithelial hepatocellular carcinoma. Am J Transl Res 8:4670–4681 Ye L, Mayerle J, Ziesch A, Reiter FP, Gerbes AL, De Toni EN (2019) The PI3K inhibitor copanlisib synergizes with sorafenib to induce cell death in hepatocellular carcinoma. Cell Death Discov 5:86. https://doi.org/10.1038/s41420-019-0165-7 Zhang Q, Dehaini D, Zhang Y, Zhou J, Chen X, Zhang L, Fang RH, Gao W, Zhang L (2018) Neutrophil membrane-coated nanoparticles inhibit synovial inflammation and alleviate joint damage in inflammatory arthritis. Nat Nanotechnol 13:1182–1190. https://doi.org/10.1038/s41565-018-0254-4 Zhang R, Gao X, Zuo J, Hu B, Yang J, Zhao J, Chen J (2020) STMN1 upregulation mediates hepatocellular carcinoma and hepatic stellate cell crosstalk to aggravate cancer by triggering the MET pathway. Cancer Sci 111:406–417. https://doi.org/10.1111/cas.14262 Zhao F, Sun X, Lu W, Xu L, Shi J, Yang S, Zhou M, Su F, Lin F, Cao F (2020) Synthesis of novel, DNA binding heterocyclic dehydroabietylamine derivatives as potential antiproliferative and apoptosis-inducing agents. Drug Deliv 27:216–227. https://doi.org/10.1080/10717544.2020.1716879 Zhou Y, Ren H, Dai B, Li J, Shang L, Huang J, Shi X (2018) Hepatocellular carcinoma-derived exosomal miRNA-21 contributes to tumor progression by converting hepatocyte stellate cells to cancer-associated fibroblasts. J Exp Clin Cancer Res 37:324. https://doi.org/10.1186/s13046-018-0965-2 Zhu AX, Rosmorduc O, Evans TRJ, Ross PJ, Santoro A, Carrilho FJ, Bruix J, Qin S, Thuluvath PJ, Llovet JM, Leberre M-A, Jensen M, Meinhardt G, Kang Y-K (2015) SEARCH: a phase III, randomized, double-blind, placebo-controlled trial of sorafenib plus erlotinib in patients with advanced hepatocellular carcinoma. J Clin Oncol 33:559–566. https://doi.org/10.1200/JCO.2013.53.7746 Zhu Y-J, Zheng B, Wang H-Y, Chen L (2017) New knowledge of the mechanisms of sorafenib resistance in liver cancer. Acta Pharmacol Sin 38:614–622. https://doi.org/10.1038/aps.2017.5 Zhu G-Q, Tang Z, Huang R, Qu W-F, Fang Y, Yang R, Tao C-Y, Gao J, Wu X-L, Sun H-X, Zhou Y-F, Song S-S, Ding Z-B, Dai Z, Zhou J, Ye D, Wu D-J, Liu W-R, Fan J, Shi Y-H (2023) CD36+ cancer-associated fibroblasts provide immunosuppressive microenvironment for hepatocellular carcinoma via secretion of macrophage migration inhibitory factor. Cell Discov 9:25. https://doi.org/10.1038/s41421-023-00529-z Zou S, Tong Q, Liu B, Huang W, Tian Y, Fu X (2020) Targeting STAT3 in cancer immunotherapy. Mol Cancer 19:145. https://doi.org/10.1186/s12943-020-01258-7