Os cientistas usam nanopartaculas de lipadios para direcionar com precisão a edição de genes para o fagado
Os pesquisadores desenvolveram nanopartaculas que carregam um pacote de edia§a£o de genes especificamente para o fígado de camundongos, reduzindo com sucesso os naveis de colesterol LDL
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)
Os pesquisadores desenvolveram nanopartaculas que carregam um pacote de edição de genes especificamente para o fígado de camundongos, reduzindo com sucesso os naveis de colesterol LDL "ruim" no sangue. Juntamente com estudos anteriores que demonstram a entrega ao cérebro ou células T, sugere uma possibilidade futura de terapias de edição de genes altamente direcionadas. Crédito: Tufts University
A tecnologia de edição de genoma CRISPR surgiu como uma nova ferramenta poderosa que pode mudar a forma como tratamos doena§as. O desafio ao alterar a genanãtica de nossas células, no entanto, écomo fazer isso de forma segura, eficaz e direcionada especificamente para o gene, tecido e órgão que precisa de tratamento. Cientistas da Tufts University e do Broad Institute of Harvard e do MIT desenvolveram nanopartaculas exclusivas compostas de lipadios - moléculas de gordura - que podem empacotar e entregar maquinaria de edição de genes especificamente para o fagado. Em um estudo publicado hoje no Proceedings of the National Academy of Sciences, eles mostraram que podem usar as nanopartaculas de lipadios (LNPs) para entregar com eficiência a maquinaria CRISPR ao fígado de camundongos, resultando na edição especafica do genoma e na redução dos naveis de colesterol no sangue em até57% - uma redução que pode durar por pelo menos vários meses com apenas uma injeção.
O problema do colesterol alto atinge mais de 29 milhões de americanos, de acordo com os Centros para Controle e Prevenção de Doena§as. A condição écomplexa e pode se originar de vários genes , bem como de escolhas nutricionais e de estilo de vida, portanto, não éfa¡cil de tratar. Os pesquisadores da Tufts and Broad, no entanto, modificaram um gene que poderia fornecer um efeito protetor contra o colesterol elevado se pudesse ser desativado pela edição do gene.
O gene no qual os pesquisadores se concentraram codifica a enzima semelhante a angiopoietina 3 (Angptl3). Essa enzima diminui a atividade de outras enzimas - lipases - que ajudam a quebrar o colesterol. Se os pesquisadores conseguirem eliminar o gene Angptl3, eles podera£o deixar as lipases fazerem seu trabalho e reduzir os naveis de colesterol no sangue. Acontece que algumas pessoas de sorte tem uma mutação natural em seu gene Angptl3, levando a naveis consistentemente baixos de trigliceradeos e colesterol de lipoproteana de baixa densidade (LDL), comumente chamado de colesterol "ruim", em sua corrente sanguínea sem quaisquer desvantagens clanicas conhecidas.
"Se pudermos replicar essa condição eliminando o gene angptl3 em outros, temos uma boa chance de ter uma solução segura e de longo prazo para o colesterol alto", disse Qiaobing Xu, professor associado de engenharia biomédica da Escola de Engenharia e autor correspondente do estudo. "Na³s apenas temos que nos certificar de que entregamos o pacote de edição de genes especificamente para o fígado para não criar efeitos colaterais indesejados."
A equipe de Xu foi capaz de fazer exatamente isso em modelos de mouse. Apa³s uma única injeção de nanopartaculas lipadicas embaladas com codificação de mRNA para CRISPR-Cas9 e um RNA de guia aºnico direcionado a Angptl3, eles observaram uma redução profunda no colesterol LDL em até57% e nos naveis de trigliceradeos em cerca de 29%, ambos os quais permaneceram nesses naveis reduzidos por pelo menos 100 dias. Os pesquisadores especulam que o efeito pode durar muito mais do que isso, talvez limitado apenas pela lenta renovação das células no fagado, que pode ocorrer ao longo de um período de cerca de um ano. A redução do colesterol e dos trigliceradeos depende da dose, então seus naveis podem ser ajustados com a injeção de menos ou mais LNPs em uma única injeção, disseram os pesquisadores.
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Em comparação, uma versão existente aprovada pela FDA de LNPs carregados com mRNA de CRISPR são poderia reduzir o colesterol LDL em no ma¡ximo 15,7% e os trigliceradeos em 16,3% quando testado em camundongos, de acordo com os pesquisadores.
O truque para fazer um LNP melhor era personalizar os componentes - as moléculas que se juntam para formar bolhas ao redor do mRNA. Os LNPs são compostos de lipadios de cadeia longa que tem uma cabea§a carregada ou polar que éatraada pela a¡gua, uma cauda de cadeia de carbono que aponta para o meio da bolha contendo a carga útil e um ligante quamico entre eles. Tambanãm estãopresentes o polietilenoglicol e, sim, atémesmo um pouco de colesterol - que tem um papel normal nas membranas lipadicas para torna¡-las menos gotejantes - para reter melhor seu conteaºdo.
Os pesquisadores descobriram que a natureza e a proporção relativa desses componentes pareciam ter efeitos profundos na entrega de mRNA ao fagado, então eles testaram os LNPs com muitas combinações de cabea§as, caudas, ligantes e proporções entre todos os componentes para sua capacidade de direcionar o fígado células. Como a potaªncia in vitro de uma formulação de LNP raramente reflete seu desempenho in vivo, eles avaliaram diretamente a especificidade e eficácia de entrega em camundongos que tem um gene repa³rter em suas células que acende em vermelho quando ocorre a edição do genoma. No final das contas, eles encontraram um LNP carregado com mRNA de CRISPR que iluminou apenas o fígado de camundongos, mostrando que poderia fornecer ferramentas de edição de genes de forma especafica e eficiente ao fígado para fazer seu trabalho.
Os LNPs foram construados a partir de trabalhos anteriores na Tufts, onde Xu e sua equipe desenvolveram LNPs com até90% de eficiência na entrega de mRNA a s células. Uma característica única dessas nanopartaculas era a presença de ligações dissulfeto entre as longas cadeias lipadicas. Fora das células, os LNPs formam uma estrutura esfanãrica esta¡vel que bloqueia seu conteaºdo. Quando estãodentro de uma canãlula, o ambiente interno quebra as ligações dissulfeto para desmontar as nanopartículas O conteaºdo éentão liberado de forma rápida e eficiente para a canãlula. Ao prevenir a perda fora da canãlula, os LNPs podem ter um rendimento muito maior na entrega de seu conteaºdo.
"CRISPR éuma das ferramentas terapaªuticas mais poderosas para o tratamento de doenças com etiologia genanãtica. Recentemente, vimos a primeira trilha clanica humana para terapia CRISPR habilitada pela entrega de LNP para ser administrada sistemicamente para editar genes dentro do corpo humano. Nosso LNP plataforma desenvolvida aqui tem grande potencial para tradução clanica ", disse Min Qiu, pesquisador de pa³s-doutorado no laboratório de Xu em Tufts. "Imaginamos que, com esta plataforma LNP em ma£os, poderaamos agora tornar o CRISPR uma abordagem prática e segura para tratar um amplo espectro de doenças ou distúrbios hepa¡ticos", disse Zachary Glass, estudante graduado no laboratório de Xu. Qiu e Glass são os co-autores do estudo.