Peptide-based drug carriers and preclinical nanomedicine applications for endometriosis treatment
Date
2023-04-13
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Abstract
Endometrioos on sage günekoloogiline haigus, mis esineb kuni 10% viljakas eas naistest. Haigust iseloomustavad väljaspool emakat kasvavad endomeetriumi ehk emaka sisemise limaskesta kolded, mis tekitavad patsientidel kroonilist kõhuvalu ja sageli ka viljatust. Kasutusel olevad endometrioosi ravi- ja diagnostikavõimalused on piiratud. Tihti ei oma farmakoloogiline ravi pikaajalist efekti ning süsteemse manustamise tõttu põhjustab see kõrvaltoimeid. Seetõttu on vajalik uute endometrioosi ravistrateegiate väljatöötamine. Paljud potentsiaalsed ravi-sihtmärgid asuvad aga tegelikkuses haiguskoldes rakkude sees. Seetõttu on mitmete haiguste puhul suurimaks väljakutseks see, et ravimid peavad kohale jõudma oma rakusiseste sihtmärk-molekulideni. Sellest tulenevalt oli antud töö eesmärgiks välja töötada või uuskasutada peptiididel põhinevaid ravimkandjaid nanomeditsiini rakenduste jaoks, keskendudes põhiliselt endometrioosile.
Alustuseks kasutati geeniekspressiooni reguleerivate lühikeste interfereeruvate RNAde (siRNA) transpordiks rakku sisenevaid peptiide ehk süstikpeptiide. Esmalt disainiti seeria uusi peptiide, mis reageerivad keskkonna füüsilistele parameetritele ja suudavad efektiivselt siRNAd rakkudesse sisse viia nii in vitro kui ka in vivo tingimustes. Teiseks testiti süstikpeptiid/siRNA nanopartiklite ravimpotentsiaali vähi ja endometrioosi eelkliinilistes rakukultuuri mudelites. Olenemata süstikpeptiidide kõrgest rakku sisenemise efektiivsusest ei oma nad koespetsiifilisust ja see võib süsteemse in vivo manustamise korral viia ravimite ebaspetsiifilise tervetesse kudedesse akumuleerumiseni. Seega uuriti viimase töö sammuna sihtmärk-kude spetsiifiliselt äratundvate peptiidide ehk kullerpeptiidide kasutusvõimalusi. Kõige edukamaks peptiidiks endometrioosi puhul osutus PL1 kullerpeptiid, millega kaetud hõbe-nanopartiklid näitasid spetsiifilist kuhjumist 2D ja 3D rakukultuurides ja seondumist kliinilistele endometrioosi koeproovidele.
Kokkuvõtvalt näitasid antud doktoritöö tulemused süstikpeptiid/siRNA nanopartiklite abil kahe haiguskudede kasvu ja progresseerumist mõjutava geeni mahasurumise sobivust endometrioosi raviks. Kombineerides nanopartikleid hormonaalse ravimiga, suurenes raviefekt veelgi. Lisaks eelnevale näitasid PL1-kaetud nanopartiklid suurt potentsiaali endometrioosi koespetsiifiliste ravimite ja diagnostikameetodite väljatöötamiseks. Kõike arvesse võttes pakuvad käesoleva doktoritöö tulemused välja mitu siirdemeditsiinilise väärtusega peptiididel põhinevat nanomeditsiini rakendust endometrioosi raviks.
Endometriosis is a frequent gynecological disease that influences up to 10% of women in reproductive age. The disorder is defined by the presence of uterus inner mucosal layer, endometrium, tissue outside the uterine cavity. The symptoms include chronic pelvic pain and often infertility. The current diagnostic and therapeutic strategies for endometriosis are limited. The treatment often lacks long-term efficacy and causes adverse effects due to systemic administration of pharmaceutics. Therefore, novel treatment strategies for endometriosis are needed. Many highly potential therapeutic targets are in fact located inside the diseased cells. Thus, the biggest challenge for successful treatment of various conditions is that the therapeutic candidates have to reach their intracellular target molecules at the intended target sites. Hence, the objective of this study was to develop or repurpose peptide-based drug carriers for nanomedical applications mainly focusing on endometriosis. To start with, highly potent drug delivery vehicles, cell-penetrating peptides (CPPs) were used as short interfering RNA (siRNA) carriers that aim to adjust the therapeutic gene expression levels. First, a series of peptides was designed that display adaptive response to the physical environment and are highly efficient in transporting siRNA into cells both in vitro and in vivo. Second, the therapeutic potential of CPP/siRNA nanoparticles was investigated in cancer and endometriosis preclinical cell culture models. Despite being very efficient in cellular internalization, most CPPs are not tissue specific and might lead to unspecific accumulation of therapeutics in healthy organs when administered in vivo. Thus, as a final step, a series of specific tissue targeting peptides was investigated. PL1 homing peptide proved to be most successful candidate for endometriosis targeting. Silver nanoparticles functionalized with synthetic PL1 peptide showed specific accumulation to 2D and 3D cell cultures and clinical endometriosis tissue samples. In conclusion, the results of this thesis proved the relevance of downregulating two genes related to growth and progression by CPP/siRNA nanoparticles for endometriosis treatment. Therapeutic effects were enhanced by the combination of nanoparticles with a hormonal drug. In addition, PL1-guided nanoparticles showed high potential in precision diagnosis and tissue specific therapy of endometriosis. All in all, the findings of this thesis propose several peptide-based nanomedical applications for the treatment of endometriosis that suggest translational relevance.
Endometriosis is a frequent gynecological disease that influences up to 10% of women in reproductive age. The disorder is defined by the presence of uterus inner mucosal layer, endometrium, tissue outside the uterine cavity. The symptoms include chronic pelvic pain and often infertility. The current diagnostic and therapeutic strategies for endometriosis are limited. The treatment often lacks long-term efficacy and causes adverse effects due to systemic administration of pharmaceutics. Therefore, novel treatment strategies for endometriosis are needed. Many highly potential therapeutic targets are in fact located inside the diseased cells. Thus, the biggest challenge for successful treatment of various conditions is that the therapeutic candidates have to reach their intracellular target molecules at the intended target sites. Hence, the objective of this study was to develop or repurpose peptide-based drug carriers for nanomedical applications mainly focusing on endometriosis. To start with, highly potent drug delivery vehicles, cell-penetrating peptides (CPPs) were used as short interfering RNA (siRNA) carriers that aim to adjust the therapeutic gene expression levels. First, a series of peptides was designed that display adaptive response to the physical environment and are highly efficient in transporting siRNA into cells both in vitro and in vivo. Second, the therapeutic potential of CPP/siRNA nanoparticles was investigated in cancer and endometriosis preclinical cell culture models. Despite being very efficient in cellular internalization, most CPPs are not tissue specific and might lead to unspecific accumulation of therapeutics in healthy organs when administered in vivo. Thus, as a final step, a series of specific tissue targeting peptides was investigated. PL1 homing peptide proved to be most successful candidate for endometriosis targeting. Silver nanoparticles functionalized with synthetic PL1 peptide showed specific accumulation to 2D and 3D cell cultures and clinical endometriosis tissue samples. In conclusion, the results of this thesis proved the relevance of downregulating two genes related to growth and progression by CPP/siRNA nanoparticles for endometriosis treatment. Therapeutic effects were enhanced by the combination of nanoparticles with a hormonal drug. In addition, PL1-guided nanoparticles showed high potential in precision diagnosis and tissue specific therapy of endometriosis. All in all, the findings of this thesis propose several peptide-based nanomedical applications for the treatment of endometriosis that suggest translational relevance.
Description
Väitekirja elektrooniline versioon ei sisalda publikatsioone
Keywords
endometriosis, cell-penetrating peptides, drug carriers, nanomedicine