Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches
Welsh J.A., Goberdhan D.C.I., O'Driscoll L., Buzas E.I., Blenkiron C., Bussolati B., Cai H., Di Vizio D., Driedonks T.A.P., Erdbrugger U., Falcon-Perez J.M., Fu Q.-L., Hill A.F., Lenassi M., Lim S.K., Mahoney M.y.G., Mohanty S., Moller A., Nieuwland R., Ochiya T., Sahoo S., Torrecilhas A.C., Zheng L., Zijlstra A., Abuelreich S., Bagabas R., Bergese P., Bridges E.M., Brucale M., Burger D., Carney R.P., Cocucci E., Crescitelli R., Hanser E., Harris A.L., Haughey N.J., Hendrix A., Ivanov A.R., Jovanovic-Talisman T., Kruh-Garcia N.A., Faustino V.K.-L., Kyburz D., Lasser C., Lennon K.M., Lotvall J., Maddox A.L., Martens-Uzunova E.S., Mizenko R.R., Newman L.A., Ridolfi A., Rohde E., Rojalin T., Rowland A., Saftics A., Sandau U.S., Saugstad J.A., Shekari F., Swift S., Ter-Ovanesyan D., Tosar J.P., Useckaite Z., Valle F., Varga Z., van der Pol E., van Herwijnen M.J.C., Wauben M.H.M., Wehman A.M., Williams S., Zendrini A., Zimmerman A.J., Thery C., Witwer K.W., Ahmad S., Ahmed D.A.K., Ahmed S.H., Aikawa E., Akbar N., Akiyoshi K., Al-Adra D.P., Al-Masawa M.E., Albanese M., Alberro A., Alcaraz M.J., Alexander-Brett J., Alexander K.L., Ali N., Alibhai F.J., Allelein S., Allenby M.C., Almeida F., de Almeida L.P., Almousa S.W., Altan-Bonnet N., Altei W.F., Alvarez-Llamas G., Alvarez C.L., An H.J., Anand K., Andaloussi S.E.L., Anderson J.D., Andriantsitohaina R., Ansari K.I., Anselmo A., Antoniou A., Aqil F., Arab T., Archer F., Arif S., Armstrong D.A., Arntz O.J., Arsene P., Arteaga-Blanco L., Asokan N., Aspelin T., Atkin-Smith G.K., Aubert D., Ayyar K.K., Azlan M., Azoidis I., Becot A., Bach J.-M., Bachurski D., Bae S., Bagge R.O., Baj-Krzyworzeka M., Balaj L., Balbi C., van Balkom B.W.M., Ballal A.R., Bano A., Banzet S., Bare Y., Barile L., Barman B., Barranco I., Barreca V., Bart G., Barteneva N.S., Basso M., Batish M., Bauer N.R., Baxter A.A., Bazie W.W., Bazzan E., Beaumont J.E.J., Bebawy M., Bebelman M.P., Bedina-Zavec A., Beetler D.J., Beke-Somfai T., Belleannee C., Benedikter B.J., Benediktsdottir B.E., Berardi A.C., Bergamelli M., Bertolini I., Bhattacharyya A., Bhattacharyya S.N., Biller S.J., Billottet C., Bissler J.J., Blanc-Brude O., Blijdorp C.J., Bobis-Wozowicz S., Bodart-Santos V., Bodnar B.R., Boilard E., Boireau W., Bokun V., Bollard S.M., Bollini S., Bongiovanni A., Bongiovanni L., Bonifay A., Boppart M.D., Borras F.E., Bosch S., Boselli D., Bottini M., Bouffard J., Boulanger C.M., Boutros P.C., Boyadjian O., Boysen A.T., Bozkurt B.T., Bramich K.P., Braun F., Del Carmen Bravo-Miana R., Breakefield X.O., Brenna S., Brennan K., Brennan M.A., Breyne K., Brigstock D.R., Brisson A.R., Brodie C., Bruno K.A., Bucci C., Buch S., Buck A., Bukva M., Bulte J.W.M., Buratta S., Burgy O., Burnier J.V., Burrows K., Busatto S., Buzas K., Byrd J.B., Caceres-Verschae A., Caires H.R., Campos-Silva C., Camussi G., de Candia P., Carceller C., Fernandez-Becerra C., Carrasco A.G.M., Carter D.R.F., Cavallaro S., Cavallero S., Cavallero S., Cerda-Troncoso C., Chahwan R., Chalupska R., Chamley L.W., Chandra P.K., Chang W.-W., Charest A., Chen C., Chen H., Chen Q., Chen S., Chen S., Chen Y., Cheng L., Chernyshev V.S., Chetty V.K., Chitti S.V., Cho S.-G., Cho Y.-K., Choi B.H., Chutipongtanate S., Cicardi M.E., Cifuentes-Rius A., Ciullo A., Clayton A., Cleary J.A., Cocozza F., Coffey R.J., Collino F., Colombo F., Colosetti P., Compan-Bertomeu A., Constanzo J., Corbeil D., Cordeiro-da-Silva A., Costa J., Couch Y., Courageux Y., Coutant K., Coyle B., Cretich M., Cronemberger-Andrade A., Crossland R.E., Cucher M.A., Czystowska-Kuzmicz M., D'Acunzo P., D'Agnano I., D'Agostino V.G., D'Arrigo D., D'Souza-Schorey C., Dagur R.S., Danielson K.M., Das S., Dauphin T., Davidson S.M., Davies O.G., Davies R.L., Davis C.N., Deep G., Degosserie J., Van Delen M., Deliwala V., Dellar E.R., Van Deun J., Dev A., Deville S., Devitt A., Dhondt B., Dieterich L.C., Dittmer D.P., Dobosh B., Dobra G., Dogra N., Dohi E., Dolo V., Domashevich T.V., Dominici M., Dong L., Dore E., Dragovic R.A., Drittanti L., Droste M., Duan W., Durmaz E., Dutta S., Eguchi T., Eichenberger R.M., Eitan E., Ekstrom K., Eldh M., Elie-Caille C., Enciso-Martinez A., Esmaeili R., Ettelaie C., Forsonits A.I., Fabbri M., Falasca M., Fan H., Fatima F., Fazeli A., Fernandez-Rhodes M., Fernandez-Prada C., Ferraro M.J., Ferreira J., Ferreira R.F., Figueroa-Hall L.K., Figueroa-Valdes A.I., Fioretti P.V., Flenady S., Flores-Bellver M., Fok E.K., Fonseka P., Forbes K., Ford V.J., Fornaguera C., Forte D., Forte S., Fortunato O., Franklin J.L., Freitas D., Frelet-Barrand A., Fujita Y., Gartner K., Gorgens A., Gabriel A.M., Gabrielli M., Gabrielsson S., Galinsoga A., Galisova A., Gamage T.K.J.B., Gao Y., Garcia-Contreras M., Garcia Garcia M.M., Garcia M.N., Gargiulo E., Garibotti H.G.-K., Mc Gee M.M., Genard G.C., Geraci F., Ghanam J., Ghatak S., Ghavami M., Ghebosu R.E., Gho Y.S., Ghosal S., Giamas G., Giebel B., Gilbert C., Gimona M., Girao H., Giusti I., Gizzie E.A., Glamoclija S., Glass S.E., Gobbo J., Godbole N., Goetz J.G., Gololobova O., Gomez-Florit M., Goncalves J.P., Gorgun C., Gori A., Gorska S., Graner M.W., Grau G.E., Grech L., Greening D.W., Gross R.M., Gross J.C., Gruber J., Gualerzi A., Guanzon D., Gudbergsson J.M., Guerin C.L., Guerra F., Guillen M.I., Gujar V., Guo W., Gupta V.B., Gupta V.K., Gustafson D., Gyukity-Sebestyen E., Holker P., Hade M.D., Hagey D.W., Han C., Han P., Hanayama R., Handberg A., Harada M., Harmati M., Harrison P., Harrison R.A., Haynes P.A., He M., Hegyesi H., van Herwijnen M.J.C., Hisey C.L., Hochberg F.H., Nolte-'tHoen E.N.M.N., Holcar M., Holder B., Holnthoner W., Holthofer H., Hooper D.C., Hosseini-Beheshti E., Hosseinkhani B., Howard J., Howe K.L., Hoyle N.R., Hrdy J., Hu G., Huang Y., Huber V., Hudoklin S., Hufnagel A., Hulett M.D., Hunt S., Hyenne V., Di Ianni E., Iannotta D., Ibrahim A.G.E., Ibrahim S.A., Ikezu S., Ikezu T., Im H., Inal J.M., Inic-Kanada A., Inngjerdingen M., Inoshima Y., Ivanova A., Izquierdo E., Jorgensen M.M., Jackson H.K., Jacobsen S., Jadue F., Javeed N., Jay S.M., Jayachandran M., Jayasinghe M.K., Jenster G., Jeppesen D.K., Jeronimo C., Jiang L., Jin J., Jingushi K., Jo D.-G., Joerger-Messerli M.S., Jones J.C., Jones M.K., de Jong O.G., Ferrante A.W., Coleman L.G., Juncker D., Jung S., Jurek B., Jurga M., Justilien V., Kabani M., Kalluri R., Kamali-Moghaddam M., Kanada M., Kang T., Kano S.-I., Kaparakis-Liaskos M., Karnas E., Karoichan A., Kashanchi F., Kashani S.A., Kashyap N.N., Katsur M., Kau-Strebinger S., Kauffman A.C., Kaur S., Kehoe O., Kelwick R.J.R., Kenari A.N., Kestecher B.M., Keulers T.G., Van Keuren-Jensen K.R., Khalaj K., Khamari D., Khanabdali R., Khomyakova E., Khoo A., Kim D.H., Kim D., Kim H.S., Kim I.-S., Kim S., Kim Y., Kima P.E., Kislinger T., Klingeborn M., Knight R., Komuro H., Koncz A., Konstantinou T., van der Koog L., Kooijmans S.A.A., Kornek M.T., Kosanovic M., Kostallari E., Koukoulis T.F., Kourembanas S., Krameralbers E.-M., Kralj-Iglic V., Krasemann S., Krasnodembskaya A.D., Krawczynska N.J., Kreft M.E., Kuehn M.J., Kuipers M.E., Kulaj K., Kuligowski J., Kumagai Y., Kumar A., Kumar S., Kumar S., Kumari M., Kundrotas G., Kurochkin I.V., Kuroda M., Kurzawa-Akanbi M., Kweskin S.J., Lazaro-Ibanez E., Lorincz A., Lai A., Lai C.P., Laitinen S., Landreville S., Lange S., Langevin S.M., Langlois M.-A., Languino L.R., Lannigan J., Lark D.S., Larregina A.T., Laurent L.C., Laurin D., Lavieu G., Lawson C., Le Lay S., Leandro K., Ledreux A., Lee C., Lee D.-S., Lee H., Lee H., Lee S.Y., Lee T.R., Lee W.-L., Lefterov I., Lei X., Leivo J., Lemaire Q., Leme A.F.P., Lemon S.M., Lenzini S., Leor J., Levy E., Li B., Li G., Li J.J., Li Q., Li X., Liang X., Lim R., Limongi T., Line A., Lins P.P., Lippens L., Liu G., Llorente A., Longjohn M.N., van de Loo F.A.J., Lorenowicz M.J., Lorico A., Loudig O., Loyer X., Lozano-Andres E., Lu B., Lu Q., Lubart Q., Lucien F., Lunavat T.R., Lundberg L.E., Lundy D.J., Luoto J.C., Lyden D.C., Muller J.A., Macphee D.J., Madec E., Magana S.M., Mahairaki V., Mahoney M.y.G., Malhi H., Malnou C.E., Mamand D.R., Man K., Manno M., Mantel P.-Y., Marangon T., Marban E., Marcilla A., Maremanda K.P., Margolis L., Marinas-Pardo L., Maric I., Martin S.S., Martinez-Martinez E., Martel C., Martin-Duque P., Martin-Jaular L., Martinez-Murillo P.A., Martinez-Pacheco S., Martins-Marques T., Mary B., Marzan A.L., Matamoros-Angles A., Mathivanan S., Matsuzaki J., Mayan M.D., Mazzeo C., Mbengue M., McCann M.J., McIlvenna L.C., McVey M.J., Meisner-Kober N., Mellergaard M., Melli G., Menck K., Menjivar N.G., Menon R., Mentkowski K.I., Miklavcic J.J., Miklosi A.G., Milutinovic B., Minciacchi V.R., Mirzaei M., Mishra S., Mitchell M.I., Mladenovic D., Mohamadi E., Momen-Heravi F., Mondal S.K., Monguio-Tortajada M., Moon J., Morandi M.I., Moreau V., Moreira L.R., Morelli A.E., Mori M.A., Morimoto M., Mosser M., Motaung T.E., Moussay E., Mugoni V., Mullier F., Muraca M., Murugesan S., Musante L., Musico A., Nemeth A., Nemeth K., Nadeau A., Nam G.-H., Naora H., Natoli R., Nawaz M., Nazarenko I., Ndukaife J.C., Nedeva C., Nejsum P., Nelissen I., Neri C., Neri T., Neviani P., Ng C.Y., van Niel G., Nikiforova N., Nimrichter L., Nitin C., Njock M.-S., Noel D., Noghero A., Nolan J.P., Noppen S., Noren Hooten N., da Silva Novaes A., O'loghlen A., Oesterreicher J., Oh S.W., Olah A., Olivier M., Ong S.L., Ortiz A., Osorio O.A., Ostergaard O., Osorio O.A., Osteikoetxea X., Ostrowski M., Otaegui D., Otahal A., Ozawa P.M.M., Ozkocak D.C., Paloczi K., Perez-Gonzalez R., Pachane B.C., Padinharayil H., Paget D., Paggetti J., Palacio P.C.L., Pallasch C.P., Palmulli R., Pang B., Paniushkina L., Pantazi P., Paolini L., Papademetrio D.L., Parisse P., Park D.J., Park J., Park Y.-G., Patton J.G., Peake N.J., Pegtel D.M., Peinado H., Perut F., Pfaffl M.W., Pfeiffer A., Phan T.K., Phinney D.G., Phylactou L.A., Picciolini S., Pietrowska M., Piffoux M., Pinheiro C., Pink R.C., Pleet M.L., Pocsfalvi G., Poh Q.H., Poojary G., Poon I.K.H., Poppa G., Del Portillo H.A., Pospichalova V., Potter S., Powell B.H., Powis S.J., Prada I., Prasadam I., Preusser C., Pua H.H., Pucci F., Puhm F., Puig B., Pulliam L., Purnianto A., Puutio J.M.M., Quilang R.C., Rabbani P.S., Rackov G., Radeghieri A., Radu C.M., Raffai R.L., Raghav A., Rahbari M., Rahman M.D.M., Rahman M.M., Rai A.J., Raimondo S., Raju S., Rak J., Ramaswamy L., Ramirezricardo J., Ramirez M.I., Rani S., Raposo G., Rather H.A., Razim A., Reale A., Reategui E., Reddel C.J., Reddy S.K., Redenti S., Reed S.L., Regevrudzki N., Reiners K.S., Resnik N., Rice G.E., Ricklefs F.L., Rilla K., Rimmer M.P., Roballo K.C.S., Robbins P.D., Roberts D.D., Roca J., Rodal A.A., Rodrigues-Junior D.M., Rodrigues M.L., Roefs M.T., Rogers R.G., Romani R., Romano M., Rome S., Romih R., Romolo A., De Rossi T., Rouschop K.M., Routenberg D.A., Roux Q., van Royen M.E., Rozo A.J., Rufino-Ramos D., Rughetti A., Russell A.E., Rutter S.F., Rysmakhanov M.S., Sanchez C.A., Sadovsky Y., Safavi-Sohi R., Sagar R., Saidu N.E.B., Saint-Pol J., Salas-Huenuleo E., Salazar-Puerta A.I., Saleem A., Salekdeh G.H., Salomon C., Salviano-Silva A., Salybekov A.A., Samuels M., Santavanond J.P., Santoro J., Santos M., Sanwlani R., Saul M.J., Schoyen T.H., Schabussova I., Scharrig E., Schekman R., Schiavi-Tritz J., Schiffelers R.M., Schmid A.M., Schneider R., Schneider S., Schoeberlein A., Schorey J.S., Seo N., Seras-Franzoso J., Shahi S., Shatnyeva O., Shea D.F., Shelke G.V., Shetty A.K., Shiba K., Shiju T.M., Shrivastava S., Shukla S., Siljander P.R.-M., Silva A.M., Singh A.P., Singh S., Skliar M., Skog J., Sluijter J.P.G., Snyder O.L., Soekmadji C., Somaida A., Somiya M., Soroczynska K., Sotillo J., Souza-Fonseca-guimaraes F., Spada S., Spiers H.V.M., Spitzberg J.D., Srivastava A., Srivastava A.K., Stepien E.L., St-Denis-bissonnette F., Stahl P.D., Stam J., Stambouli O., Stanton B.A., Stassen F.R.M., Staufer O., Steiner L., Stepanova G., Stoka V., Stoorvogel W., von Strandmann E.P., Strunk D., Stylli S.S., Su H., Subramanian S., Sui B., Sukreet S., Sulaiman E., Sung B.H., Sunkara V., Suo Z., Svenningsen P., Swatler J., Symonds E.K.C., Szeifert V., Szigyarto I.C., Toth E.A., Tasli N.P., Tahara H., Takahashi R.-U., Takakura Y., Takikawa O., Takov K., Tang V.A., Taverna S., Tawil N., Teeuwen L., Tejedor S., Tertel T., Thakur A., Thompson-Felix T., Tian C., Tikhonov A., Tiwari S., Toh W.S., Tomes J.J., Tonoli E., Trinidad C.V., Tritten L., Trivedi R., Troyer Z., Tsamchoe M., Tscherrig V., Tsering T., Turkova K., Tutanov O.S., Ueda K., Upadhya D., Urabe F., Urbanelli L., Urzi O., Vacchi E., Vader P., Vago R., Valadi H., Valkonen S., Varas-Godoy M., Varga Z., Vasconcelos M.H., Vechetti I.J., Veiga S.I., Vella L.J., Velot E., Verweij F.J., Vestad B., Vinay L., Viola M., Visnovitz T., Vreeland W.N., Vukman K.V., Wade P.K., van de Wakker S.I., Walther L., Wang T., Wang X., Watson D.C., Weaver A.M., Webber J.P., Weber V., Weiss L., Weiss M.L., Weiss R., Weissleder R., Wen Y., de Wever O., Wheelock A.M., White K.E., Whitehead B., Whiteside T.L., Whitley J., Wiener Z., Van Wijnen A.J., Wiklander O.P.B., Winston C.N., Wolf M., Wolfram J., Wu L., Wu Y., Wysmolek M.E., Xander P., Xavier C.P.R., Xiao Y., Xu R., Yanez-Mo M., Yamamoto T., Yamamoto Y., Yamamoto Y., Yan X., Yang L., Yang Y., Yarani R., Yea K., Yedigaryan L., Yenuganti V.R., Yerneni S.S., Yeung V., Yildizhan Y., Yin H., Yokoi A., Yoshioka Y., You Y., Yuan L.-Q., Yunga S.T., Zakeri A., Zani A., Zanoni M., Zappulli V., Zarovni N., Zarubova J., Zempleni J., Zekas V., Zhang H., Zhang Q., Zhao Z., Zhou Y., Zickler A.M., Zimmermann P., Zivkovic A.M., Zocco D., Zuba-Surma E.K., Zubair H.
Article, Journal of Extracellular Vesicles, 2024, DOI Link
View abstract ⏷
Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its ‘Minimal Information for Studies of Extracellular Vesicles’, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.
Exosomes in the visual system: New avenues in ocular diseases
Manukonda R., Attem J., Yenuganti V.R., Kaliki S., Vemuganti G.K.
Article, Tumor Biology, 2022, DOI Link
View abstract ⏷
Exosomes are a subgroup of membrane-bound extracellular vesicles secreted by all cell types and present virtually in all biological fluids. The composition of exosomes in the same cell type varies in healthy and disease conditions. Hence, exosomes research is a prime focus area for clinical research in cancer and numerous age-related metabolic syndromes. Functions of exosomes include crucial cell-To-cell communication that mediates complex cellular processes, such as antigen presentation, stem cell differentiation, and angiogenesis. However, very few studies reported the presence and role of exosomes in normal physiological and pathological conditions of specialized ocular tissues of the eye and ocular cancers. The eye being a protected sense organ with unique connectivity with the rest of the body through the blood and natural passages, we believe that the role of exosomes in ocular tissues will significantly improve our understanding of ocular diseases and their interactions with the rest of the body. We present a review that highlights the existence and function of exosomes in various ocular tissues, their role in the progression of some of the neoplastic and non-neoplastic conditions of the eyes.
Drug Loading and Functional Efficacy of Cow, Buffalo, and Goat Milk-Derived Exosomes: A Comparative Study
Ahmed F., Tamma M., Pathigadapa U., Reddanna P., Yenuganti V.R.
Article, Molecular Pharmaceutics, 2022, DOI Link
View abstract ⏷
Quite recently, milk exosomes have been recognized as efficient drug delivery systems owing to their biocompatibility and easy availability for scale-up technologies. However, there are no reports of comparative studies with regards to drug delivery by milk exosomes derived from different species. In this study, we isolated and characterized milk exosomes of cow, buffalo, and goat by various techniques and tried to understand their drug loading capacity and functional efficiency in HepG2, HCT116, and A549 cells by using doxorubicin. Doxorubicin was loaded to milk exosomes by three methods, that is, incubation, saponin treatment, and sonication. The isolated exosomes were found to be spherical with a size of <200 nm and displayed specific markers, namely, CD81, HSP70, HSC70, and miRNAs. Drug loading studies revealed that goat milk exosomes had the highest loading capacity across all three methods. Doxorubicin-encapsulated goat milk exosomes resulted in the inhibition of cell viability, with low IC50values in HepG2, HCT-116, and A549 cells. Doxorubicin-encapsulated goat exosomes displayed better IC50values than cow and buffalo milk-derived counterparts. In line with this, the ability of doxorubicin-encapsulated goat milk exosomes to induce apoptosis in HepG2 and HCT-116 cells was higher than that of cow and buffalo milk exosomes and free doxorubicin. Furthermore, unbound goat milk exosomes significantly reduced cell viability as compared to cow and buffalo milk exosomes. The transepithelial transport assay shows that doxorubicin-loaded milk exosomes transport doxorubicin efficiently as compared to free doxorubicin in vitro. Doxorubicin released from milk exosomes shows a biphasic release pattern, burst release followed by sustained release. These observations are important in light of the emerging importance of milk-derived exosomes as drug carriers to treat cancers.
Comprehensive Analysis of Serum Small Extracellular Vesicles-Derived Coding and Non-Coding RNAs from Retinoblastoma Patients for Identifying Regulatory Interactions
Manukonda R., Yenuganti V.R., Nagar N., Dholaniya P.S., Malpotra S., Attem J., Reddy M.M., Jakati S., Mishra D.K., Reddanna P., Poluri K.M., Vemuganti G.K., Kaliki S.
Article, Cancers, 2022, DOI Link
View abstract ⏷
The present study employed nanoparticle tracking analysis, transmission electron microscopy, immunoblotting, RNA sequencing, and quantitative real-time PCR validation to characterize serum-derived small extracellular vesicles (sEVs) from RB patients and age-matched controls. Bioinformatics methods were used to analyze functions, and regulatory interactions between coding and non-coding (nc) sEVs RNAs. The results revealed that the isolated sEVs are round-shaped with a size < 150 nm, 5.3 × 1011 ± 8.1 particles/mL, and zeta potential of 11.1 to −15.8 mV, and expressed exosome markers CD9, CD81, and TSG101. A total of 6514 differentially expressed (DE) mRNAs, 123 DE miRNAs, and 3634 DE lncRNAs were detected. Both miRNA-mRNA and lncRNA-miRNA-mRNA network analysis revealed that the cell cycle-specific genes including CDKNI1A, CCND1, c-MYC, and HIF1A are regulated by hub ncRNAs MALAT1, AFAP1-AS1, miR145, 101, and 16-5p. Protein-protein interaction network analysis showed that eye-related DE mRNAs are involved in rod cell differentiation, cone cell development, and retinol metabolism. In conclusion, our study provides a comprehensive overview of the RB sEV RNAs and regulatory interactions between them.
Milk exosomes elicit a potent anti-viral activity against dengue virus
Yenuganti V.R., Afroz S., Khan R.A., Bharadwaj C., Nabariya D.K., Nayak N., Subbiah M., Chintala K., Banerjee S., Reddanna P., Khan N.
Article, Journal of Nanobiotechnology, 2022, DOI Link
View abstract ⏷
Background: Exosomes are nano-sized vesicles secreted by various cells into the intra and extracellular space and hence is an integral part of biological fluids including milk. In the last few decades, many research groups have proved the potential of milk exosomes as a sustainable, economical and non-immunogenic drug delivery and therapeutic agent against different pathological conditions. However, its anti-viral properties still remain to be unearthed. Methods: Here, we have been able to isolate, purify and characterize the milk derived exosomes from Cow (CME) and Goat (GME) and further studied its antiviral properties against Dengue virus (DENV), Newcastle Disease Virus strain Komarov (NDV-K) and Human Immunodeficiency Virus (HIV-1) using an in-vitro infection system. Results: TEM, NTA and DLS analysis validated the appropriate size of the isolated cow and goat milk exosomes (30–150 nm). Real-time PCR and immunoblotting results confirmed the presence of several milk exosomal miRNAs and protein markers. Our findings suggest that GME significantly decreased the infectivity of DENV. In addition, we confirmed that GME significantly reduces DENV replication and reduced the secretion of mature virions. Furthermore, heat inactivation of GME did not show any inhibition on DENV infection, replication, and secretion of mature virions. RNase treatment of GME abrogates the anti-viral properties indicating direct role of exosomes in DENV inhibition. In addition GME inhibited the infectivity of NDV-K, but not HIV-1, suggesting that the GME mediated antiviral activity might be virus specific. Conclusion: This study demonstrates the anti-viral properties of milk exosomes and opens new avenues for the development of exosome-based therapies to treat viral diseases. Graphical Abstract: [Figure not available: see fulltext.]
In vitro evaluation of anticancer effects of different probiotic strains on HCT-116 cell line
Yenuganti V.R., Yadala R., Azad R., Singh S., Chiluka V., Ahire J., Reddanna P.
Article, Journal of Applied Microbiology, 2021, DOI Link
View abstract ⏷
Aim: Since the evolution of man, microbes are associated with humans, playing a vital role in the maintenance of good health. However, an imbalance in the gut microbial ecosystem is associated with several diseases including colorectal cancer (CRC). The supplementation with probiotics has been proven to be beneficial in improving CRC. In this study, we have evaluated the anticancer effects of 11 probiotic strains on human colorectal carcinoma cell line (HCT-116). Methods and Results: In this study, HCT-116 cells were treated with various concentrations (0·5, 5, 10, 20 and 200 million CFU per ml) of probiotic strains. The viability was analysed using a MTT assay and IC50 values were determined. Besides this, we evaluated the expression of multiple genes involved in the apoptosis and stress tolerance by real-time PCR. Lactobacillus reuteri (UBLRu-87), Saccharomyces boulardii (Unique-28), Bacillus clausii (UBBC-07), Bacillus coagulans (Unique-IS2), Streptococcus salivarius (UBSS-01), Lactobacillus fermentum (UBLF-31), Lactobacillus salivarius (UBLS-22), Bifidobacterium bifidum (UBBB-55) and Lactobacillus plantarum (UBLP-40) exhibited potent cytotoxicity on HCT 116 cells. Furthermore, UBLF-31 and Unique-28 induced the expression of CJUN, CFOS and CASP-9, and downregulated the expression of BCL6. UBLRu-87 and UBBB-55 induced the expression of CJUN, CFOS and CASP-9 but not BCL-6. UBLP-40, UBBC-07, UBLS-22, and Unique-IS2 induced the expression of CJUN and CASP-9 and downregulated the expression of BCL-6. Conclusion: These studies indicate the anticancer effects of selected probiotic strains by inducing apoptosis. Significance and Impact of the Study: The probiotic strains with the anticancer effects identified in this study can be proposed as potential candidates in the treatment of CRCs.
Correction to: Genome wide effects of oleic acid on cultured bovine granulosa cells: evidence for the activation of pathways favoring folliculo-luteal transition (BMC Genomics, (2021), 22, 1, (486), 10.1186/s12864-021-07817-6)
Erratum, BMC Genomics, 2021, DOI Link
View abstract ⏷
Following publication of the original article [1], it was reported that affiliations 1 and 3 were erroneously reversed. The affiliations have been correctly assigned for Vengala Rao Yenuganti and Jens Vanselow in this Correction article and the original article has been updated.
Genome wide effects of oleic acid on cultured bovine granulosa cells: evidence for the activation of pathways favoring folliculo-luteal transition
Article, BMC Genomics, 2021, DOI Link
View abstract ⏷
Background: Metabolic stress, as negative energy balance on one hand or obesity on the other hand can lead to increased levels of free fatty acids in the plasma and follicular fluid of animals and humans. In an earlier study, we showed that increased oleic acid (OA) concentrations affected the function of cultured bovine granulosa cells (GCs). Here, we focus on genome wide effects of increased OA concentrations. Results: Our data showed that 413 genes were affected, of which 197 were down- and 216 up-regulated. Specifically, the expression of FSH-regulated functional key genes, CCND2, LHCGR, INHA and CYP19A1 and 17-β-estradiol (E2) production were reduced by OA treatment, whereas the expression of the fatty acid transporter CD36 was increased and the morphology of the cells was changed due to lipid droplet accumulation. Bioinformatic analysis revealed that associated pathways of the putative upstream regulators “FSH” and “Cg (choriogonadotropin)” were inhibited and activated, respectively. Down-regulated genes are over-represented in GO terms “reproductive structure/system development”, “ovulation cycle process”, and “(positive) regulation of gonadotropin secretion”, whereas up-regulated genes are involved in “circulatory system development”, “vasculature development”, “angiogenesis” or “extracellular matrix/structure organization”. Conclusions: From these data we conclude that besides inhibiting GC functionality, increased OA levels seemingly promote angiogenesis and tissue remodelling, thus suggestively initiating a premature fulliculo-luteal transition. In vivo this may lead to impeded folliculogenesis and ovulation, and cause sub-fertility.
Exosomes: The protagonists in the tale of colorectal cancer?
Review, Biochimica et Biophysica Acta - Reviews on Cancer, 2020, DOI Link
View abstract ⏷
Exosomes, which facilitate intercellular communication, antigen presentation and shuttling of biological agents, were initially thought as the cell's garbage cargo but today, after about 40 years of their discovery, we are now beginning to understand their potential role in diagnosis and therapy of several diseases including cancers. Various studies over the decades have signified the role of exosomes in different stages of cancer. Exosomes play a key role in colorectal cancer initiation (CRC), promotion of anti- apoptotic signaling pathways, regulating tumor microenvironment, enhancing tumorigenicity, promotion of angiogenesis, stem cell proliferation and endothelial cell migration, establishment of immune suppressive environment, formation of pre- metastatic niche and metastasis. Exosomes also elicits drug resistance. Since, they have the ability to cross the biological barrier, exosomes are now being explored as an efficient target specific drug delivery system that facilitates the shipping of different biomolecules and therapeutic drugs. However, cautious and strong investigative approaches are required before approving exosomes as therapeutics or drug delivery systems. In this review, we summarize the role of exosomes in different stages of CRC and also elaborate on the applications of exosomes in diagnosis and therapy with respect to CRC.
Cultured bovine granulosa cells rapidly lose important features of their identity and functionality but partially recover under long-term culture conditions
Article, Cell and Tissue Research, 2017, DOI Link
View abstract ⏷
Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.
Fermented milk in protection against inflammatory mechanisms in obesity
Pothuraju R., Yenuganti V.R., Hussain S.A., Sharma M.
Book chapter, Immunity and Inflammation in Health and Disease: Emerging Roles of Nutraceuticals and Functional Foods in Immune Support, 2017, DOI Link
View abstract ⏷
Recent studies indicate that obesity is linked with several metabolic complications. The cluster of metabolic diseases contributes towards low grade inflammation which leads to metabolic endotoxemia. Metabolic disorders in animals and humans that follow high fat/cholesterol supplementation are associated with alteration of gut microbiota (dysbiosis). Thus diet plays an important role in the management of obesity via maintenance of proper gut microflora. Fermented milks are considered to be functional foods in which lactobacilli drive the fermentation process. The various health benefits of fermented milks are due to the presence of bioactive peptides, branched-chain amino acids and angiotensin-converting enzyme inhibitors. Here we discuss the health promoting effects of fermented milk by different lactobacilli species with a focus on adiposity and inflammatory mechanisms involved in obesity. Further, we discuss the antiobesity effect of milk-fermented dairy products (kefir and yogurt) in animal and clinical studies.
miRNAs: The hidden bioactive component of milk
Rani P., Yenuganti V.R., Shandilya S., Onteru S.K., Singh D.
Review, Trends in Food Science and Technology, 2017, DOI Link
View abstract ⏷
Background miRNAs are the non-coding ribonucleic acid species around 22 nucleotides in length that are involved in various biological processes via regulating gene expression. They are abundantly present in the milk and have been identified in the various fractions of milk. Scope and approach Milk miRNAs are emerging as the new bioactive component of milk. miRNAs have been identified in whey, lipid as well as the exosomal fraction of milk of different species. But they are majorly present as encapsulated in the lipid bilayered nano-vesicles called exosomes, which confers stability and functionality to the milk miRNAs. They are stable even under harsh conditions, including high temperature, enzymatic treatments and low pH, indicating their physiological significance. Most of the abundant milk miRNAs are having immune-related functions, inclining the attention towards the role of milk miRNAs in providing immunity as well as nutrition. These milk exosomes not only participate in the intercellular communication but also, facilitate encapsulating cellular component including miRNAs to perform cellular functions. Overall, the stability of exosomes along with their potential to communicate with the target cells strengthens the view of encapsulated milk miRNAs as a bioactive component of milk. Key findings and conclusions Milk miRNAs are stable against degradative conditions, may be bioavailable, may enter into the blood circulation and may affect the gene expression of target genes in recipient cells, influencing the immunity of infants and adult consumer of bovine milk. However, more studies are needed to be done to discover their physiological stability and bioactive potential.
Oleic acid induces down-regulation of the granulosa cell identity marker FOXL2, and up-regulation of the Sertoli cell marker SOX9 in bovine granulosa cells
Article, Reproductive Biology and Endocrinology, 2017, DOI Link
View abstract ⏷
During negative energy balance, the concentration of different fatty acids, especially of oleic acid (OA) increases in the follicular fluid of cattle. Previously, we showed that OA induced morphological, physiological and molecular changes in cultured bovine granulosa cells. In our present study we analyzed effects of OA on the expression of markers for granulosa and Sertoli cell identity, FOXL2 and SOX9, respectively, in addition to effects on the FOXL2 regulated genes ESR2, FST, PTGS2 and PPARG. The results showed that OA down-regulated FOXL2, ESR2, FST and PPARG but up-regulated PTGS2 and SOX9. From these data we conclude that OA can compromise granulosa cell functionality and may initiate trans-differentiation processes in bovine granulosa cells. This novel mechanism may be causally involved in postpartum fertility problems of lactating dairy cows.
Endotoxin induced TLR4 signaling downregulates CYP19A1 expression through CEBPB in buffalo granulosa cells
Article, Toxicology in Vitro, 2017, DOI Link
View abstract ⏷
Estrogen is essential for growth and development of ovarian follicles. Infections associated with E. coli or Endotoxin (LPS) suppress estradiol production by the downregulation of CYP19A1 expression. However, the molecular mechanism of its down regulation is not yet known. To elucidate the molecular mechanisms of LPS-mediated downregulation of CYP19A1 gene expression, we studied the effect of LPS and TLR4 signaling pathway inhibitor (OxPAPC, OxPAPC-Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine) on CYP19A1 expression, and expression of CEBPB and CEBPB binding on CYP19A1 proximal promoter (CYP19A1 PII) in buffalo granulosa cells in vitro. The results showed that LPS (10.25 μg/ml) significantly declined the expression of CYP19A1 gene. In further experiments, inhibitor studies confirmed the involvement of TLR4 in LPS induced CYP19A1 gene down regulation in buffalo granulosa cells. LPS promoted higher levels of CEBPB at cellular and nuclear level in granulosa cells. Chromatin immunoprecipitation results showed, that LPS induces higher amount of CEBPB binding on the CYP19A1 PII. Further, TLR4 inhibitor attenuated the LPS induced implications. In conclusion, our results demonstrated that CEBPB could be a potential regulator for LPS mediated downregulation of CYP19A1 and decline of 17-beta estradiol levels in buffalo granulosa cells.
Oleic acid induces specific alterations in the morphology, gene expression and steroid hormone production of cultured bovine granulosa cells
Article, General and Comparative Endocrinology, 2016, DOI Link
View abstract ⏷
After parturition, one of the major problems related to nutritional management that is faced by the majority of dairy cows is negative energy balance (NEB). During NEB, excessive lipid mobilization takes place and hence the levels of free fatty acids, among them oleic acid, increase in the blood, but also in the follicular fluid. This accumulation can be associated with serious metabolic and reproductive disorders. In the present study, we analyzed the effects of physiological concentrations of oleic acid on cell morphology, apoptosis, necrosis, proliferation and steroid production, and on the abundance of selected transcripts in cultured bovine granulosa cells. Increasing oleic acid concentrations induced intracellular lipid droplet accumulation, thus resulting in a foam cell-like morphology, but had no effects on apoptosis, necrosis or proliferation. Oleic acid also significantly reduced the transcript abundance of the gonadotropin hormone receptors, FSHR and LHCGR, steroidogenic genes STAR, CYP11A1, HSD3B1 and CYP19A1, the cell cycle regulator CCND2, but not of the proliferation marker PCNA. In addition, treatment increased the transcript levels of the fatty acid transporters CD36 and SLC27A1, and decreased the production of 17-beta-estradiol and progesterone. From these data it can be concluded that oleic acid specifically affects morphological and physiological features and gene expression levels thus altering the functionality of granulosa cells. Suggestively, these effects might be partly due to the reduced expression of FSHR and thus the reduced responsiveness to FSH stimulation.
The gene expression pattern induced by high plating density in cultured bovine and buffalo granulosa cells might be regulated by specific miRNA species
Yenuganti V.R., Baddela V.S., Baufeld A., Singh D., Vanselow J.
Article, Journal of Reproduction and Development, 2015, DOI Link
View abstract ⏷
Precise regulation of cell type-specific gene expression profiles precedes the profound morphological reorganization of somatic cell layers during folliculogenesis, ovulation and luteinization. Cell culture models are essential to the study of corresponding molecular mechanisms of gene regulation. In a recent study, it was shown that an increased cell plating density can largely change gene expression profiles of cultured bovine granulosa cells. In our present study, we comparatively analyzed cell plating density effects on cultured bovine and buffalo granulosa cells. Cells were isolated from small- to medium-sized follicles (2–6 mm) and cultured under serum-free conditions at different plating densities. The abundance of selected marker transcripts and associated miRNA candidates was determined by quantitative real-time RT-PCR. We found in both species that the abundance of CYP19A1, CCNE1 and PCNA transcripts was remarkably lower at a high plating density, whereas VNN2 and RGS2 transcripts significantly increased. In contrast, putative regulators of CYP19A1, miR-378, miR-106a and let-7f were significantly higher in both species or only in buffalo, respectively. Also miR-15a, a regulator of CCNE1, was upregulated in both species. Thus, increased plating density induced similar changes of mRNA and miRNA expression in granulosa cells from buffalo and cattle. From these data, we conclude that specific miRNA species might be involved in the observed densityinduced gene regulation.
Suitable housekeeping genes for normalization of transcript abundance analysis by real-time RT-PCR in cultured bovine granulosa cells during hypoxia and differential cell plating density
Baddela V.S., Baufeld A., Yenuganti V.R., Vanselow J., Singh D.
Article, Reproductive Biology and Endocrinology, 2014, DOI Link
View abstract ⏷
Background: Bovine granulosa cell culture models are important to understand molecular mechanisms of ovarian function. Folliculogenesis and luteinization are associated with increasing density of cells and local hypoxic conditions. The current study identified two reliable housekeeping genes useful for gene normalization in granulosa cells under different in vitro conditions. Methods: During the current experiments cells were subjected to different biological and physical stimuli, follicle stimulating hormone, different initial cell plating density and hypoxia. Transcript abundance of seven housekeeping genes was quantified by real-time RT-PCR with co-amplification of the respective external standard. Results: Three of the genes, GAPDH, HMBS, and HPRT1 were found to be regulated by initial cell plating density, five of them, GAPDH, HMBS, HPRT1, RPLP0 and RPS18 under hypoxic conditions, but none of them after FSH stimulation. In detail, GAPDH was up regulated, but HPRT1 and HMBS were down regulated at high density and under hypoxia. Expression of RPLP0 and RPS18 was inconsistent, but was significantly down-regulated in particular at high cell density combined with hypoxia. In contrast, TBP and B2M genes were neither regulated under different plating density conditions nor by hypoxia as they showed similar expression levels under all conditions analyzed. Conclusions: The present data indicate that TBP and B2M are appropriate housekeeping genes for normalization of transcript abundance measured by real-time RT-PCR in granulosa cells subjected to different plating densities, oxygen concentrations and FSH stimulation.
Conjugated linoleic acids attenuate LPS-induced pro-inflammatory gene expression by inhibiting the NF-κB translocation through PPARγ in buffalo granulosa cells
Article, American journal of reproductive immunology (New York, N.Y. : 1989), 2014, DOI Link
View abstract ⏷
PROBLEM: In granulosa cells, TLR4-mediated LPS-induced immune response interferes with ovarian granulosa cell function. METHOD OF STUDY: LPS-induced pro-inflammatory gene expression was monitored by real-time PCR and NF-κB was determined by the immuno-blotting and immuno-staining in granulosa cells in vitro. RESULTS: The LPS (1 μg/mL) increased pro-inflammatory gene expression which was reverted back by treatment with TLR4 signaling inhibitor. Cotreatment of CLA (10 μm) with LPS did not show any effect on LPS-induced pro-inflammatory gene expression but granulosa cells pre-treated with CLA for 24 hr, attenuated LPS-induced pro-inflammatory gene expression and nuclear NF-κB. GW9662, a PPARγ-inhibitor, further increased the expression of pro-inflammatory genes. CONCLUSION: The present findings reiterated that pre-treatment with CLA can prevent LPS-induced granulosa cells dysfunction.
Attenuation of LPS induced proinplammatory gene expression by conjugated linoleic acid (CLA) in buffalo granulosa cells
Conference paper, Buffalo Bulletin, 2013,
View abstract ⏷
Lipopolysaccharide (LPS) is a component of gram negative bacterial (E. coli) outer membrane. Previous studies showed that LPS present in plasma and follicular fluid of postpartum uterine infected animals and inducing the inflammatory response in bovine granulosa cells. Conjugated linoleic acid (CLA) is a dietary fatty acid that has unique properties like antiinflammatory, anti-cancer, anti-atherogenic and anti-diabetic effects. In present study we studied the effects of LPS on proinflammatory gene expression and CLA as anti-inflammatory agent to attenuate LPS induced inflammatory gene expression in cultured buffalo granulosa cells under serum free media. Granulosa cells were cultured and treated with LPS (1.0 μg/ml) for 6 h at different time intervals and pro-inflammatory cytokines (TNFα, IL-1β and IL-6) were analyzed using qRT- PCR. Result shown that all the three pro-inflammatory cytokines were significantly (P≤0.01) up regulated within 2h and then decreased sharply and reached to almost normal level within 6 h. The Conjugated linoleic acid (CLA), ligand of PPARγ, has been shown to be a potent anti-inflammatory agent. To test, whether CLA can prevent the LPS induced pro-inflammatory cytokines expression in granulosa cells, cells were cultured 48 hrs for basal growth, after basal growth cells were co-treated with CLA (10 μM) and LPS (1.0 μg/ml) for 2 h followed by analysis of expression of pro-inflammatory cytokines. Surprisingly, CLA had no effect on pro-inflammatory cytokine expression. Further, cells were pre-treated with CLA for 24hr after basal growth and then cells were exposed to LPS for 2 h. Result showed that cells pre-treated with CLA showed significantly (P≤0.01) less expression of pro inflammatory cytokines when comparison to LPS alone. In conclusion, the results of the present study showed that CLA pre-treatment can prevent the LPS induced pro-inflammatory cytokines expression in granulosa cells in vitro. (Work supported by Department of Biotechnology (DBT), Govt. of India.).
