Respiratory disease causes an immense worldwide health burden. Nearly 25 million people suffer from end-stage lung disease in the United States alone, with a staggering ~400,000 patients dying each year. We focused on two different lung diseases: acute respiratory distress syndrome (ARDS) and a genetic mutation in alveolar type II (ATII) cells. The most affected area of the lung in these diseases is the air sacs (alveoli) and, more in specific, the epithelium. ARDS is clinically defined by severe lung damage with inflammation, and dysfunctional lungs that are collapsed or filled with fluid. Many ARDS survivors develop lung fibrosis and end-stage lung disease. On the other hand, when ATII cells are injured or diseased, the alveoli cannot function properly and therefore they cannot deliver sufficient oxygen to the bloodstream or remove carbon dioxide, causing respiratory symptoms. At this time, ameliorating the consequences of these diseases remains challenging and lung transplantation, the only definitive treatment for these patients, remains hampered by donor organ shortages, long-term rejection, and the need for immunosuppressive therapy. Final aim of this thesis work has been to find new therapeutic approaches, able to remove only the injured or defective epithelial cells, while preserving all the surrounding lung cells and architecture, in order to repair lung tissue and foster lung regeneration. 1) In vivo de-epithelialization in a rat model of ARDS. After having established an ARDS model in vivo and the feasibility and efficiency of the de-epithelialization treatment in healthy animals, CHAPS-based solution was used in order to remove the injured epithelium in the ARDS animal, maintaining the vasculature and the lung architecture intact, and to stimulate and recruit lung progenitors to repopulate the treated organ. 2) Targeted therapy for defective ATII cells disease. Our novel drug, a bioengineered therapeutic protein that joins a substance naturally produced by the lung together with the catalytic domain a toxin, that induces cell apoptosis, has been tested in vitro and in vivo on mice in order to remove only the defective cells and create a favorable environment for cells transplantation and lung regeneration.
Le malattie respiratorie gravano sui costi della sanità in tutto il mondo. Quasi 25 milioni di pazienti soffrono di malattie polmonari croniche allo stadio terminale solo negli Stati Uniti e di questi circa 400 000 muoiono ogni anno. In questo lavoro di tesi ci siamo focalizzati su due diverse malattie polmonari: la sindrome da distress respiratorio (ARDS, Acute Respiratory Distress Syndrome) e una mutazione genetica delle cellule alveolare di tipo II (ATII, Alveolar type II cells). L'area più colpita del polmone in queste malattie è l’alveolo e, più in particolare, l'epitelio alveolare. ARDS è una forma di lesione polmonare acuta, classificata come malattia infiammatoria, in cui i polmoni sono collassati o presentano edema. Molti pazienti con ARDS sviluppano, superata la fase acuta, uno stato grave di fibrosi polmonare. Inoltre, quando le cellule ATII sono geneticamente danneggiate, gli alveoli non possono funzionare correttamente e non garantiscono gli scambi gassosi, non supportando il fabbisogno di ossigeno dell’organismo e causando sintomi patologici respiratori. Attualmente, non vi sono terapie in grado di migliorare le conseguenze di queste malattie ed il trapianto di polmone, l'unico trattamento definitivo per questi pazienti, rimane ostacolato dalla carenza di organi donatori, dal possibile rigetto e dalla necessità di terapia immunosoppressiva. L'obiettivo finale di questa tesi è stato quello di trovare nuovi approcci terapeutici, in grado di rimuovere solo le cellule epiteliali lesionate o difettose, preservando tutte le altre componenti polmonari e l'architettura circostante, al fine di riparare il tessuto e favorire la rigenerazione polmonare. 1. De-epitelializzazione in vivo di un modello di ratto in cui viene indotta l’ARDS. Dopo aver dimostrato di sia di poter indurre la malattia in vivo e che di poter trattare efficientemente con la de-epitelializzazione animali sani, la soluzione a base di CHAPS per rimuovere l'epitelio è stata utilizzata su ratti malati di ARDS, mantenendo intatta la rete vascolare e l'architettura polmonare, per stimolare e reclutare i progenitori polmonari per ripopolare l'organo trattato. 1. Targetizzazione specifica di pneumociti di tipo 2 affetti da modifiche genetiche. Il nostro nuovo farmaco, una proteina bioingegnerizzata composta dall’unione di una sostanza prodotta naturalmente nel polmone con il dominio catalitico di una tossina, che induce l'apoptosi cellulare, è stato testato in vitro e in vivo su topi al fine di rimuovere solo le cellule difettose e creare un ambiente favorevole per il trapianto di cellule esogene sane e la rigenerazione polmonare.
Efficient and targeted de-epithelialization as novel therapeutic strategy for lung regeneration
PREDELLA, CAMILLA
2018/2019
Abstract
Respiratory disease causes an immense worldwide health burden. Nearly 25 million people suffer from end-stage lung disease in the United States alone, with a staggering ~400,000 patients dying each year. We focused on two different lung diseases: acute respiratory distress syndrome (ARDS) and a genetic mutation in alveolar type II (ATII) cells. The most affected area of the lung in these diseases is the air sacs (alveoli) and, more in specific, the epithelium. ARDS is clinically defined by severe lung damage with inflammation, and dysfunctional lungs that are collapsed or filled with fluid. Many ARDS survivors develop lung fibrosis and end-stage lung disease. On the other hand, when ATII cells are injured or diseased, the alveoli cannot function properly and therefore they cannot deliver sufficient oxygen to the bloodstream or remove carbon dioxide, causing respiratory symptoms. At this time, ameliorating the consequences of these diseases remains challenging and lung transplantation, the only definitive treatment for these patients, remains hampered by donor organ shortages, long-term rejection, and the need for immunosuppressive therapy. Final aim of this thesis work has been to find new therapeutic approaches, able to remove only the injured or defective epithelial cells, while preserving all the surrounding lung cells and architecture, in order to repair lung tissue and foster lung regeneration. 1) In vivo de-epithelialization in a rat model of ARDS. After having established an ARDS model in vivo and the feasibility and efficiency of the de-epithelialization treatment in healthy animals, CHAPS-based solution was used in order to remove the injured epithelium in the ARDS animal, maintaining the vasculature and the lung architecture intact, and to stimulate and recruit lung progenitors to repopulate the treated organ. 2) Targeted therapy for defective ATII cells disease. Our novel drug, a bioengineered therapeutic protein that joins a substance naturally produced by the lung together with the catalytic domain a toxin, that induces cell apoptosis, has been tested in vitro and in vivo on mice in order to remove only the defective cells and create a favorable environment for cells transplantation and lung regeneration.File | Dimensione | Formato | |
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https://hdl.handle.net/10589/151144