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Endosomal escape of protein nanoparticles engineered through humanized histidine-rich peptides

Science China Materials 2020年第4期63卷 644-653页

作者： Hector Lopez-Laguna Rafael Cubarsi Ugutz Unzueta Ramon Mangues esther vazquez Antonio Villaverde Institut de Biotecnologia i de Biomedicina Universitat Aut6noma de BarcelonaBellaterra08193 BarcelonaSpain Departament de Genetica i de Microbiologia Universitat Aut6noma de BarcelonaBellaterra08193 BarcelonaSpain CIBER de Bioingenieria Biomateriales y Nanomedicina(CIBER-BBN)C/Monforte de Lemos 3-528029 MadridSpain Departament de Matematiques Universitat Politecnica de Catalunya08034BarcelonaSpain Institut d'Investigacions Biomediques Sant Pau Hospital de la Santa Creu i Sant Pau08025 BarcelonaSpain

Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags,protein-assembling agents and endosomal-escape *** pleiotropic properties of such peptides make them appealing to design protein-based smart materials or nanoparticles for imaging or drug delivery to be produced in form of recombinant ***,the clinical applicability of H6-tagged proteins is restricted by the potential immunogenicity of these *** this study,we have explored several humanized histidine-rich peptides in tumor-targeted modular proteins,which can specifically bind and be internalized by the target cells through the tumoral marker *** were particularly interested in exploring how protein purification,self-assembling and endosomal escape perform in proteins containing the variant histidine-rich *** the tested candidates,the peptide H5 E(HEHEHEHEH)is promising as a good promoter of endosomal escape of the associated fulllength protein upon endosomal *** numerical modelling of cell penetration and endosomal escape of the tested proteins has revealed a negative relationship between the amount of protein internalized into target cells and the efficiency of cytoplasmic *** fact demonstrates that the His-mediated,proton sponge-based endosomal escape saturates at moderate amounts of internalized protein,a fact that might be critical for the design of protein materials for cytosolic molecular delivery.

关键词： protein materials nanoparticles genetic design endosomal escape poly-histidines

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Controlling self-assembling and tumor cell-targeting of protein-only nanoparticles through modular protein engineering

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Science China Materials 2020年第1期63卷 147-156页

作者： Eric Volta-Duran Olivia Cano-Garrido Naroa Serna Hector Lopez-Laguna Laura Sanchez-Garcia Mireia Pesarrodona Alejandro Sanchez-Chardi Ramon Mangues Antonio Villaverde esther vazquez Ugutz Unzueta Institut de Biotecnologia i de Biomedicina Universitat Autònoma de BarcelonaBellaterra08193 BarcelonaSpain Departament de Genetica i de Microbiologia Universitat Autònoma de BarcelonaBellaterra08193 BarcelonaSpain CIBER de Bioingeniería Biomateriales y Nanomedicina(CIBER-BBN)C/Monforte de Lemos 3-528029 MadridSpain Nanoligent SL Edifici EUREKAUniversitat Autonoma de BarcelonaBellaterra08193 BarcelonaSpain Institut dInvestigacions Biomèdiques Sant Pau and Josep Carreras Research Institute Hospital de la Santa Creu i Sant Pau08041 BarcelonaSpain Departament de Biologia Evolutiva Ecologia i Ciencies AmbientalsFacultat de BiologiaUniversitat de BarcelonaAv.Diagonal 64308028 BarcelonaSpain Institute for Research in Biomedicine(IRB Barcelona) The Barcelona Institute of Science and Technology08028 BarcelonaSpain

Modular protein engineering is suited to recruit complex and multiple functionalities in single-chain polypeptides. Although still unexplored in a systematic way, it is anticipated that the positioning of functional domains would impact and refine these activities, including the ability to organize as supramolecular entities and to generate multifunctional protein materials. To explore this concept, we have repositioned functional segments in the modular protein T22-GFP-H6 and characterized the resulting alternative fusions. In T22-GFP-H6, the combination of T22 and H6 promotes selfassembling as regular nanoparticles and selective binding and internalization of this material in CXCR4-overexpressing tumor cells, making them appealing as vehicles for selective drug delivery. The results show that the pleiotropic activities are dramatically affected in module-swapped constructs, proving the need of a carboxy terminal positioning of H6 for protein self-assembling, and the accommodation of T22 at the amino terminus as a requisite for CXCR4^+ cell binding and internalization. Furthermore, the failure of self-assembling as regular oligomers reduces cellular penetrability of the fusions while keeping the specificity of the T22-CXCR4 *** these data instruct how multifunctional nanoscale protein carriers can be designed for smart, protein-driven drug delivery, not only for the treatment of CXCR4^+ human neoplasias, but also for the development of anti-HIV drugs and other pathologies in which CXCR4 is a relevant homing marker.

关键词： nanoparticles protein materials recombinant proteins drug delivery self-assembling cancer cell targeting

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Engineering a recombinant chlorotoxin as celltargeted cytotoxic nanoparticles

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Science China Materials 2019年第6期62卷 892-898页

作者： Raquel Díaz Laura Sánchez-García Naroa Serna Alejandro Sánchez-Chardi Olivia Cano-Garrido Julieta M.Sánchez Ugutz Unzueta esther vazquez Antonio Villaverde Institut de Biotecnologia i de Biomedicina Universitat Autònoma de Barcelona Departament de Genètica i de Microbiologia Universitat Autònoma de Barcelona CIBER de Bioingeniería Biomateriales y Nanomedicina(CIBER-BBN) Servei de Microscòpia Universitat Autònoma de Barcelona Biomedical Research Institute Sant Pau(IIB-Sant Pau)and Josep Carreras Research Institute Hospital de la Santa Creu i Sant Pau Universidad Nacional de Córdoba Facultad de Ciencias ExactasFísicas y Naturales.ICTA and Departamento de QuímicaCátedra de Química Biológica.Córdoba.Argentina.CONICETInstituto de Investigaciones Biológicas y Tecnológicas(IIBYT)

Cytotoxic proteins have a wide applicability in human therapies, especially in those conditions that require efficient and selective cell killing, such as cancer. Chlorotoxin (CTX) is a small (4 kDa) basic peptide from the venom of the yellow scorpion Leiurus quinquestriatus , which blocks small-conductance chloride channels thus paralyzing the scorpion prey. Being not extremely potent as a cytotoxin (for instance when compared with ribosome-inactivating proteins), it has gained interest as a targeting agent.

关键词：重组DNA技术细胞毒活性纳米颗粒蝎毒素癌细胞可生物降解氯药物载体

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New therapeutic approaches to metastatic gastroenteropancreatic neuroendocrine tumors:A glimpse into the future

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World Journal of Gastrointestinal Oncology 2017年第1期9卷 4-20页

作者： esther Una Cidon Department of Medical Oncology Royal Bournemouth HospitalBournemouthDorset BH5 2AZUnited Kingdom

Neuroendocrine(NE) gastroenteropancreatic tumors are a heterogeneous group of neoplasias arising from neuroendocrine cells of the embryological gut. Their incidence have increased significantly over the past 3 decades probably due to the improvements in imaging and diagnosis. The recent advances in molecular biology have translated into an expansion of therapeutic approaches to these patients. Somatostatin analogs, which initially were approved for control of hormonal syndromes, have recently been proven to inhibit tumor growth. Several new drugs such as antiangiogenics and others targeting mammalian target of rapamycin pathways have been approved to treat progressive pancreatic neuroendocrine tumors(NETs) although their role in nonpancreatic is still controversial. The treatment of NETs requires a coordinated multidisciplinary approach. The management of localized NETs primarily involves surgical resection followed by surveillance. However, the treatment of unresectable and/or metastatic disease may involve a combination of surgical resection, systemic therapy, and liver-directed therapies with the goal of alleviating symptoms of peptide release and controlling tumor growth. This article will review the current therapeutic strategies for metastatic gastroenteropancreatic NETs and will take a glimpse into the future approaches.

关键词： Gastroenteropancreatic neuroendocrine tumors Peptide receptor radionuclide therapy Somatostatin analogs Octreotide Transarterial chemoembolization Carcinoid syndrome Setotonin Chromogranin

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Vasoactive intestinal peptide secreting tumour:An overview

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World Journal of Gastrointestinal Oncology 2022年第4期14卷 808-819页

作者： esther Una Cidon Department of Medical Oncology University Hospitals DorsetBournemouth BH77DWDorsetUnited Kingdom

Vasoactive intestinal peptide(VIP)secreting tumour(VIPoma)is a rare functional neuroendocrine tumour that typically arises from pancreatic islet *** present as sporadic,solitary pancreatic neoplasias with an estimated incidence of one in ten million individuals per *** around 5%of VIPomas are associated with multiple endocrine neoplasia type I *** VIP secretion produces a clinical syndrome characterized by refractory watery diarrhoea,hypokalemia and metabolic *** coupled with elevated plasma levels of VIP are *** majority of VIPomas are malignant and have already metastasized at the time of diagnosis(60%).Metastases occur most frequently in the liver,or regional lymph nodes,lungs,kidneys and *** reports of skin metastases have been *** surgical resection continues to be the only potentially curative ***,when the neoplasia cannot be excised completely,surgical debulking may provide palliative *** palliative options have included recently the peptide receptor radionuclide therapy which has shown to be effective and *** article will review all aspects of pancreatic VIPomas highlighting aspects such as clinical presentation,diagnosis and management.

关键词： VIPoma Octreotide Secretory diarrhoea Multiple endocrine neoplasia Vasoactive intestinal peptide Vermer-Morrison syndrome

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Apoptosis in skeletal muscle and its relevance to atrophy

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World Journal of Gastroenterology 2006年第46期12卷 7463-7466页

作者： esther E Dupont-Versteegden University of Arkansas for Medical Sciences Departments of Geriatrics and Physiology and Biophysics 4301 West Markham #807 Little Rock AR 72205 and University of Kentucky Department of Rehabilitation Sciences Division of Physical Therapy 900 S Limestone Lexington KY 40536-0200 United States

Apoptosis is necessary for maintaining the integrity of proliferative tissues, such as epithelial cells of the gastrointestinal system. The role of apoptosis in post mitotic tissues, such as skeletal muscle, is less well defined. Apoptosis during muscle atrophy occurs in both myonuclei and other muscle cell types. Apoptosis of myonuclei likely contributes to the loss of muscle mass, but the mechanisms underlying this process are largely unknown. Caspase-dependent as well as -independent pathways have been implicated and the mode by which atrophy is induced likely determines the apoptotic mechanisms that are utilized. It remains to be determined whether a decrease in apoptosis will alleviate atrophy and distinct research strategies may be required for different causes of skeletal muscle loss.

关键词： Skeletal muscle Apoptosis Disuse atrophy Aging

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Small mammal diversity of Mt. Kenya based oncarnivore fecal and surface bone remains

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Zoological Research 2019年第1期40卷 61-69页

作者： Ogeto Mwebi esther Nguta Veronica Onduso Ben Nyakundi Xue-Long Jiang esther N.Kioko Osteology Section Department of Zoology National Museums of Kenya Kunming Institute of Zoology Chinese Academy of Sciences Sino-African Joint Research Center Chinese Academy of Sciences

Ecological dynamics and faunal diversity documentation is normally conducted by direct observation and trapping of live animals. However,surveys of carnivore scat prey and surface bone remains, which are relatively inexpensive, can provide complementary data that expand carnivore diet breadth and may improve accuracy regarding inferences of the ecological dynamics of a given ecosystem. We used this inexpensive method to document species diversity variation with elevation on the leeward(Sirimon) and windward(Chogoria)areas of Mt. Kenya. Bone and fecal specimens were opportunistically collected by walking 2 km in opposite directions from transect points selected at 200-m intervals along the elevational gradient of the study areas. We collected a total of 220 carnivore fecal and owl pellet specimens from both study sites, which were mainly deposited by the spotted hyena(Crocuta crocuta), leopard(Panthera pardus),serval(Leptailurus serval), genet(Genetta sp.), and Mackinder's Cape owl(Bubo capensis mackinderi).Serval scats were the most common, followed by those of the spotted hyena. Scats and bones were found at the lowest density at the lowest elevations,peaked at mid-higher elevations, and then declined at the highest elevations. Based on skeletal analysis only, there were more species in Sirimon(19) than in Chogoria(12). Small fauna(rodents to duiker size bovids) formed the bulk of the identified remains,representing 87.9% of the Sirimon fauna and 90.9% of the Chogoria fauna. The genus Otomys was the dominant prey of the owl and serval in both sites. Three giraffe teeth were found at 3 500 m a.s.l. in Chogoria on the edge of Lake El is, suggesting that it is an occasional visitor to such high elevations. This study underscores the value of fecal and bone surveys in understanding the diet and diversity of mammals in ecological ecosystems,but such surveys should be complemented with analysis of hairs found in scats to obtain a more complete list of carnivore prey at M

关键词： Ecological dynamics Faunal diversity Scats Pellets Mt.Kenya

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New approaches in angiogenic targeting for colorectal cancer

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World Journal of Gastroenterology 2007年第44期13卷 5857-5866页

作者： Aleix Prat esther Casado Javier Cortés Department of Medical Oncology Vall d'Hebron University Hospital Barcelona Spain Department of Medical Oncology Vall d'Hebron University Hospital Barcelona Spain Department of Medical Oncology Hospital General de Manresa Barcelona Spain

Colorectal carcinoma （CRC） is one of the leading causes of cancer death worldwide. In the last decade, the addition of irinotecan and oxaliplatin to standard fluorouracil-based chemotherapy regimens have set the new benchmark of survival for patients with metastatic CRC at approximately 20 too. Despite these advances in the management of CRC, there is a strong medical need for more effective and well-tolerated therapies. The dependence of tumor growth and metastasis on blood vessels makes angiogenesis a rational target for therapy. One of the major pathways involved in this process is the vascular endothelial growth factor （VEGF） and its receptors （VEGFR）. In 2004, the first agent targeting angiogenesis, bevacizumab （BV）, was approved as an adjunct to first-line cytotoxic treatment of metastatic CRC. The role of BV as part of adjuvant treatment and in combination with other targeted therapies is the subject of ongoing trials. However, BV is associated with an increase in the risk of arterial thromboembolic events, hypertension and gastrointestinal perforations and its use must be cautious. Novel VEGFR TK inhibitors with different ranges of nanomolar potencies, selectivities, and pharmacokinetic properties are entering phase 111 trials for the treatment of cancer. Conversely, one of these novel agents, vatalanib, has been shown not to confer survival benefit in first and second-line treatment of advanced CRC. The basis of these findings is being extensively evaluated. Ongoing and new well-designed trials will define the optimal clinical application of the actual antiangiogenic agents, and, on the other hand, intensive efforts in basic research will identify new agents with different antiangiogenic approaches for the treatment of CRC. In this review we discuss and highlight current and future approaches in angiogenic targeting for CRC.

关键词： Angiogenesis inhibitors Vascular endothelial growth factor VEGF receptors Bevacizumab Vatalanib Colorectal carcinoma

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Unexplained liver laceration after metastasis radiofrequency ablation

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World Journal of Gastroenterology 2009年第40期15卷 5103-5105页

作者： esther Ua Javier Trueba Jose Manuel Montes Medical Oncology Service Clinical University Hospital of Valladolid Radiology Service Clinical University Hospital of Valladolid

Many studies have established the role of radiofrequency (RF) ablation as a minimally invasive treatment for liver metastases. Although relatively safe, several complications have been reported with the increased use of RF ablation. We describe here a case of unexplained liver laceration after a RF procedure. A woman who presented a solitary metachronous liver metastasis underwent RF ablation treatment for this lesion. Six hours later the patient displayed fatigue and pallor. Emergency blood tests showed a haemoglobin level of < 7 g/dL and markedly elevated transaminase levels. A computed tomography examination revealed two areas of liver laceration with haematoma, one of them following the path of the needle and the other leading away from the f irst. Following a blood transfusion, the patient was haemodynamically stable and completely recovered 24 h later. The patient remained in bed for 1 wk. No surgical intervention was required, and she was discharged 1 wk later.

关键词： Colon cancer Liver haemorrhage Liver laceration Liver metastases Radiofrequency ablation

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Systemic treatment of hepatocellular carcinoma:Past,present and future

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World Journal of Hepatology 2017年第18期9卷 797-807页

作者： esther Una Cidon Department of Medical Oncology Royal Bournemouth HospitalBournemouth BH7 7DWUnited Kingdom

Hepatocellular carcinoma(HCC) is a common neoplasia which represents the second leading cause of cancer related death. Most cases occur in developing countries, but its incidence is rising in Western countries due to hepatitis C. Although hepatitis therapies have evolved and the HCC screening has increased in several areas, 40% present with advanced disease which is only amenable for palliative systemic treatment. HCC continues posing a challenge, in part due to the inherent chemoresistance of this neoplasia, the pharmacologic challenges due to an ill liver, difficulty in assessing radiological responses accurately, etc. Traditional chemotherapy have shown some responses without clear survival benefit, however, sorafenib demonstrated advantages in survival in advanced HCC when liver function is kept and recently immunotherapy seems to be a promising approach for some patients. This article will briefly expose the most relevant systemic treatment modalities to offer a general view from the past to the future.

关键词： Hepatocellular carcinoma Alphafetoprotein Sorafenib Nivolumab MEK

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