O DESAFIO<\/p>\n
A terapia de captura de neutr\u00f5es (NCT) \u00e9 uma forma altamente precisa de radioterapia, que explora a capacidade de alguns is\u00f3topos espec\u00edficos gerarem part\u00edculas de elevada energia (LET) eficientes na irradica\u00e7\u00e3o de c\u00e9lulas cancer\u00edgenas. A recente instala\u00e7\u00e3o de aceleradores de \u00faltima gera\u00e7\u00e3o em ambiente hospitalar, potencializou a NCT como uma op\u00e7\u00e3o apelativa no tratamento de cancros altamente agressivos (ex. c\u00e9rebro, pele e cabe\u00e7a). No entanto, os f\u00e1rmacos atuais s\u00e3o ainda bastante limitados no seu desempenho, incluindo baixa estabilidade qu\u00edmica, toxicidade sist\u00eamica, baixa seletividade e persist\u00eancia intracelular durante a irradia\u00e7\u00e3o de neutr\u00f5es, causando efeitos secund\u00e1rios graves.<\/p>\n
O NOSSO OBJETIVO<\/p>\n
CarboNCT apresenta uma abordagem completamente inovadora para o desenvolvimento de agentes nanoterapeuticos multifuncionais mais eficientes para NCT. Para o efeito, ser\u00e1 explorado o novo conceito de nanoc\u00e1psulas de carbono (CNCs), capazes de acomodar elevadas concentra\u00e7\u00f5es de nucl\u00eddeos ativos 6Li, 157\/155Gd e 10B na sua cavidade interna. Neste sentido, ser\u00e3o exploradas pela primeira vez nanocapsulas baseadas em \u201ccarbon nanohorns\u201d (CNHs) e pontos qu\u00e2nticos de carbono (CNDs). A implementa\u00e7\u00e3o de modelos computacionais at\u00f3micos ajustados aos dados experimentais permitir\u00e1 atingir otimizar DENSIDADE DE ESP\u00c9CIES ATIVAS DE NCT NA CAVIDADE INTERNA das CNCs. \u00c9 importante salientar que as esp\u00e9cies de NCT estar\u00e3o localizadas na cavidade interna das CNCs, EVITANDO TOXICIDADE E DEGRADA\u00c7\u00c3O. A superf\u00edcie das CNCs ser\u00e1 modificada atrav\u00e9s da fabrica\u00e7\u00e3o microflu\u00eddica de revestimento de hidrogel biotivo, carregado com o f\u00e1rmaco\u00a0clinicamente aprovado indocianina verde-doxorrubicina (DOX). O controle das caracter\u00edsticas f\u00edsico-qu\u00edmicas do revestimento de hidrogel atrav\u00e9s do sistema microflu\u00eddico, permitir\u00e1 modular comportamento fisiol\u00f3gico das CNCs tal como\u00a0providenciar a liberta\u00e7ao controlada do agente de quimioterapia (dox). adicionalmente, a superf\u00edcie das CNCs ser\u00e1 conjugada com ligandos espec\u00edficos que reconhecem receptores expressos nas c\u00e9lulas cancer\u00edgenas de forma a incrementar a internaliza\u00e7\u00e3o celular e a acumula\u00e7\u00e3o e difus\u00e3o\u00a0tumoral.<\/p>\n
Os CNCs MULTIFUNCIONAIS desenvolvidos funcionar\u00e3o como um nanoplataformas espec\u00edficas capazes de fornecer seletivamente uma combina\u00e7\u00e3o terap\u00eautica (nct \/quimio), possibilitando simultaneamente a sua monitoriza\u00e7\u00e3o por bioimagem de alta resolu\u00e7\u00e3o. As CNCs apresentam fluoresc\u00eancia intr\u00ednseca (CNDs e indocianina-DOX ), possibilitando assim a monitoriza\u00e7\u00e3o em tempo real da biodistribui\u00e7\u00e3o semiquantitativa por an\u00e1lise confocal.(Santos et al. 2020) Por forma a melhor compreender a intera\u00e7\u00e3o da terapia \u00e0 base de CNCs multifuncionais e o microambiente complexo do tecido tumoral, ser\u00e3o realizados estudos usando modelos multicelulares de tumores da cabe\u00e7a produzidos por bioimpress\u00e3o 3D. Estes modelos preditivos de tumor 3D in-vitro, possibilitar\u00e3o a dete\u00e7\u00e3o e monitoriza\u00e7\u00e3o in-situ da terapia multimodal baseada em CNCs de forma mais realista. Esta informa\u00e7\u00e3o ser\u00e1 cr\u00edtica para a triagem do diagn\u00f3stico terap\u00eautico e a otimiza\u00e7\u00e3o da dosagem de irradia\u00e7\u00e3o de neutr\u00f5es (tempo\/flu\u00eancia), por forma a implementar tratamentos pr\u00e9-cl\u00ednicos altamente eficientes de NCT.<\/p>\n
AS NOSSAS COMPETENCIAS COMPLEMENTARES PARA ATINGIR O OBJETIVO<\/p>\n
CarboNCT \u00e9 uma proposta de investiga\u00e7\u00e3o cient\u00edfica multidisciplinar desenvolvida por um cons\u00f3rcio de tr\u00eas unidades de investiga\u00e7\u00e3o com compet\u00eancias complementares capazes de criar sinergias que capitalizem e otimizem os recursos existentes no sentido de atingir os principais objetivos do projeto:<\/p>\n
1) TEMA-UA, s\u00edntese e caracteriza\u00e7\u00e3o de novas CNCs preenchidas com agentes ativos de NCT.<\/p>\n
2) CICECO-UA, modula\u00e7\u00e3o computacional aplicada para otimiza\u00e7\u00e3o da s\u00edntese qu\u00edmica das CNCs e funcionaliza\u00e7\u00e3o superficial com hidrogel bioativo.<\/p>\n
3) iCBR-UC e UA-TEM, Avalia\u00e7\u00e3o de biocompatibilidade e desenvolvimento de modelos terap\u00eauticos preditivos atrav\u00e9s tumores multicelulares obtidos por bioimpress\u00e3o 3D.<\/p>\n
As equipas de investiga\u00e7\u00e3o envolvidas no CarboNCT apresentam elevadas compet\u00eancias e experi\u00eancia na \u00e1rea, evidenciadas pela coordena\u00e7\u00e3o de v\u00e1rios projetos Nacionais\/Europeus e publica\u00e7\u00e3o de in\u00fameros artigos cient\u00edficos e patentes, que \u00e9 complementada por uma estreita colabora\u00e7\u00e3o do PI com o Laborat\u00f3rio de Energia Nuclear Aplicada (LENA),\u00a0com elevados conhecimentos na aplica\u00e7\u00e3o cl\u00ednica de NCT. O cons\u00f3rcio ser\u00e1, portanto, capaz de gerar conhecimento significativo para o desenvolvimento de novos agentes nanoterap\u00eauticos de NCT para poss\u00edvel aplica\u00e7\u00e3o cl\u00ednica.<\/p>\n
THE CHALLENGE<\/p>\n
Neutron capture therapy (NCT) is a highly precise form of radiotherapy, that exploits specific isotopes to produce high linear energy transfer (LET) particles that can cause cancer cell death. The recent installation of accelerator in hospitals enviorment, advanced NCT clinical practice for treating highly aggressive cancers, (Koivunoro et al. 2019) namely high-grade gliomas, primaries or cerebral metastases of melanoma, and head\/neck cancers, where other conventional therapies were ine\ufb00ective. However, current pharmaceuticals present substantial limitations for successful clinical translation including, low chemical stability, selectivity, and persistence intra-cellularly during neutron irradiation, causing severe side e\ufb00ects.<\/p>\n
OUR GOAL<\/p>\n
CarboNCT presents an innovative approach to designing more efficient multifunctional nanotherapeutic NCT agents by exploring 10B, 157\/155Gd, 6Li active nuclides. For that purpose, we will explore\u00a0the new concept of carbon nanocapsules (CNCs), able to accommodate high concentrations of active nuclides in their internal cavity. Here, carbon nanohorns (CNHs) and carbon nanodots (CNDs) will be investigated\u00a0for the first time for the synthesis of CNCs Computer models will be applied to elucidate the strength and specificity of the interaction between the load and the di\ufb00erent nanocarriers for improved filling yields. the high density of nct active species in the inner cavity of the nanocarrier will provide the possibility to implement a new disruptive NCT nanotherapy. Significantly, the NCT species will be located in the inner cavity of the CNCs avoiding toxicity and degradation.<\/p>\n
The CNCs external surfaces will be coated with bioactive hydrogel loaded with clinically approved chemotherapeutic indocyanine green-doxorubicin (dox) by microfluidics-based fabrication. The precise control over the physicochemical features of the hydrogel<\/p>\n
coating by the microfluidic system allows modulating the CNCs physiological behaviour as well as controlling\u00a0drug release profile (DOX). Furthermore, biopolymer will be conjugated with ligands that recognize receptors that are highly expressed in cancer cells or the tumor microenvironment (TME), increasing CNCs cellular uptake, tumor targeting and di\ufb00usion. The multifunctional CNCs will operate as selectively delivering nanoagents for an e\ufb00ective chemo-neutron therapy combination monitored by high-resolution bioimaging, while avoiding systemic side e\ufb00ects typical of conventional cancer therapies. CNCs present intrinsic fluorescence (CNDs and tagged DOX), that can provide relevant information regarding the real-time monitorization of the\u00a0biodistribution by confocal analysis, following the vision for the 21st century of personalized medicine. To better understand the interplay between the complex<\/p>\n
TME\u00a0and the delivery of the CNCs-based therapy, a head and neck tumor model will be mimicked by using 3D-bioprinted multicellular tumors. The predictive in-vitro 3D tumor models will allow the in-situ confocal bio-detection and monitorization of CNCs-based therapy to uncover the underlying transduction pathways involved in reducing malignant tumor progression. This information is critical for the therapeutic diagnosis screening and the optimization of the neutron irradiation dosage (time\/fluency), to conduct highly e\ufb00icient NCT pre-clinical tests.<\/p>\n
OUR COMPLEMENTARY COMPETENCIES TO ACHIEVE THE GOAL<\/p>\n
CarboNCT is a multidisciplinary scientific research proposal formed by a consortium of three research units with complementary expertise to create synergies that capitalize and optimize existing means and resources to reach the main specific goals of the project:<\/p>\n
1) TEMA-UA, synthesis and characterization of novel CNCs filled with NCT active agents.<\/p>\n
2) CICECO-UA, applied computer modelling to support the chemical synthesis, and controlled bioactive hydrogel coating of CNCs.<\/p>\n
3) ICBR-UC and TEMA-UA, In vitro biocompatibility analysis of CNCs and the development of predictive models using bioprinted 3D multicellular head\/neck tumor.<\/p>\n
The research teams involved in CarboNCT have high competencies and experience in the field, evidenced by the coordination of several National\/European projects and publication of numerous scientific articles and patents, that is complemented by a strict collaboration of the PI with the Laboratory for Applied Nuclear Energy (LENA), with high expertise in the medical application of NCT. Therefore, the consortium will be able to generate critical knowledge for the development of novel NCT nanomedicines towards clinical application.<\/p>\n