The transformation from normal to malignant phenotype in human cancers is associated with aberrant cell-surface glycosylation. Mucin 1 (MUC1), the heavily glycosylated cell-surface mucin, is altered in both, expression and glycosylation pattern in many cancers. The presence of truncated glycan structures, often capped by sialic acid, commonly known as tumor-associated carbohydrate antigens (TACAs), play key roles in tumor initiations, progression, and metastasis. Accumulating evidence suggests that expression of TACAs is associated with escape of immune defenses. Human macrophage galactose-type lectin (hMGL, HML, CD301 or CLEC10A), a C-type lectin expressed by antigen presenting cells (APC), is a receptor of mucin-type TACAs, -GalNAc (Thomsen nouvelle antigen; Tn; CD175) and its 2,6-sialylated derivative (sTn; CD175s). To date, the relative contributions of these glycans, as well as underlying peptide backbone, and different degrees of valency, on binding thermodynamics and kinetics with hMGL remains elusive. In order to discern the subtle utility of these distinct features, chemical syntheses of the MUC1, HGVTSAPDTRPAPGSTAPPA tandem repeat sequence, and its site-specific serine (Ser) and threonine (Thr) glycosylated analogs were carried out. Circular dichroism (CD) spectroscopy experiments detected increasing structural order of the Thr glycopeptides compared to its nonglycosylated analogs. Isothermal titration calorimetry (ITC) data analysis of lectin binding to the Thr glycopeptides invariably showed enthalpy-driven processes. Affinity enhancement of the Thr glycopeptides for hMGL occurred relative to free GalNAc, revealing an increasing trend in affinity by one order of magnitude, for mono- (KD = 6-8 μM) to triglycosylated (KD = 600 nM) MUC1 peptides. To delineate the relevance of the solvent structure in the protein carbohydrate recognition process, experiments in D2O were performed, exposing enthalpy-entropy compensation differences. KinITC analysis highlighted prolonged complex lifetimes. Furthermore, atomic force microscopy (AFM) based dynamic single-molecule force spectroscopy (SMFS) provided molecular level insight into the energy landscapes governing recognition of the MUC1(Tn)-hMGL complexes. In summary, our results suggest that contact with hMGL critically depends on the type of TACA, nature of the vicinity surrounding the glycan, and its density. This highlights the importance and current efforts in design of prophylactic and therapeutic cancer vaccines with special emphasis on the synthetic glycopeptide vaccines. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2021. / FAU Electronic Theses and Dissertations Collection

O objetivo deste estudo foi descrever e mapear a toxicidade local e sistêmica da aplicação da vacina de DNA HSP 65 do Mycobacterium leprae em pacientes com formas avançadas de carcinoma epidermóide de cabeça e pescoço e definir qual a dose máxima tolerada. Trata-se de um estudo prospectivo, randomizado, aberto, sem grupo controle, de fase 1, com utilização de uma nova vacina para o tratamento de dezoito pacientes com carcinoma epidermóide avançado de cabeça e pescoço, sem opção terapêutica curativa, índice de Karnofsky maior que 70%, sem outra doença sistêmica grave. Propôs-se 3 grupos de 6 indivíduos, cada grupo recebendo diferente dose da vacina, respectivamente 150ug; 600ug e 1200ug por dose. A administração desta foi feita em 3 injeções com intervalo de 21 dias. Durante 90 dias os pacientes eram rigorosamente avaliados clinico e laboratorialmente quanto à ocorrência de eventos adversos (EA). A pesquisa de EA foi baseada no Common Terminology Criteria for Adverse Events (CTCAE) elaborado pelo National Cancer Institute (NCI). No primeiro grupo (150ug) um paciente morreu por sangramento abundante de úlcera tumoral antes do fim do protocolo. Todos os pacientes referiram aumento da dor; três tiveram maior edema e dois piora da astenia; três tiveram infecções de pele e/ou tecido subcutâneo da cabeça e pescoço e um infecção do trato respiratório superior. No 2º grupo (600ug) três pacientes faleceram antes dos 90 dias de protocolo por causas não relacionadas ao tratamento, descritas a seguir: complicações de gastrostomia, rápida progressão tumoral e hemorragia fatal e carcinomatose pulmonar. Neste grupo um paciente não teve EA; cinco tiveram piora da dor; quatro aumento do edema; dois maior astenia; três celulite de face e dois apresentaram sinusite aguda. Como neste grupo observou-se toxicidade classificada como relacionada ao tratamento graus 3 e 4 em mais de um terço dos pacientes decidiu-se escalonar para baixo a dose no 3º grupo para 400ug. No último grupo três pacientes morreram por progressão tumoral rápida, sem completar o protocolo, então incluiu-se três novos indivíduos. Todos tiveram maior dor; três aumento do edema; três piora da astenia; um caso de erisipela; três com infecção pulmonar e um com sinusite aguda; um paciente apresentou linfonodomegalia peri-tumoral após receber a vacina. Concluiu-se que a administração desta vacina neste grupo de pacientes é segura na dose de 400ug e tem como principal toxicidade aumento dos sinais inflamatórios na lesão tumoral e maior ocorrência de infecções locais e respiratórias. / This study goal was to describe and graduate the local and systemic toxicity of intratumoral injections of HSP (Heat Shock Protein) 65 DNA vaccine in advanced-stage head and neck squamous cell carcinoma (SCCHN) patients and to define the highest well tolerated dose for them. This is a prospective, non-randomized, uncontrolled, phase 1 study, using a new vaccine to treat eighteen patients with advanced-stage SCCHN patients without any option for curative treatment, Karnofsky performance status greater than 70% and no organ failure. The patients were divided into 3 groups of 6 patients each one, receiving different vaccine doses, 150ug; 600ug e 1200ug per dose, respectively. They received three injections with a 21 days interval. The patients were rigorously evaluated into their clinical and laboratory aspects looking for adverse events (AE) during 90 days. Toxic effects were monitored according to National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE). In the first group (150ug) one patient died due to an ulcerate lesion extensive bleeding before the protocol end. All the patients referred pain increase; tree had greater edema; two had strong fatigue; tree presented with cutaneous infections of head and neck and one with an acute sinusitis. In the second group (600ug) three patients died before the 90th day protocol, all considered unrelated to treatment, following described: gastrostomy complications, a fatal bleeding after rapid progression of the tumor and pulmonary carcinomatosis. In this group one patient didn\'t have adverse events; five had pain worsening; four had increase of edema; two had greater fatigue; tree had facial cellulitis and two had acute sinusitis. Due to grade 3 and 4 adverse events occurred in more than one third of patients of this group we decided to lower the dose of the third group to 400ug. In the last group three patients also died before protocol completion, all due to cancer progression, so we had to include three more patients in this group. All of them presented greater pain; tree had increasing of edema; tree had fatigue worsening; tree had pulmonary infections; one had acute sinusitis and lymphoadenomegalia besides the tumor lesion. We concluded that intratumoral injections of M.leprae HSP 65 DNA in advanced-stage SCCHN patients is safe at the dose of 400ug/injection and cause tumor flare adverse events and an increase rate of local and respiratory infections.

O objetivo deste estudo foi descrever e mapear a toxicidade local e sistêmica da aplicação da vacina de DNA HSP 65 do Mycobacterium leprae em pacientes com formas avançadas de carcinoma epidermóide de cabeça e pescoço e definir qual a dose máxima tolerada. Trata-se de um estudo prospectivo, randomizado, aberto, sem grupo controle, de fase 1, com utilização de uma nova vacina para o tratamento de dezoito pacientes com carcinoma epidermóide avançado de cabeça e pescoço, sem opção terapêutica curativa, índice de Karnofsky maior que 70%, sem outra doença sistêmica grave. Propôs-se 3 grupos de 6 indivíduos, cada grupo recebendo diferente dose da vacina, respectivamente 150ug; 600ug e 1200ug por dose. A administração desta foi feita em 3 injeções com intervalo de 21 dias. Durante 90 dias os pacientes eram rigorosamente avaliados clinico e laboratorialmente quanto à ocorrência de eventos adversos (EA). A pesquisa de EA foi baseada no Common Terminology Criteria for Adverse Events (CTCAE) elaborado pelo National Cancer Institute (NCI). No primeiro grupo (150ug) um paciente morreu por sangramento abundante de úlcera tumoral antes do fim do protocolo. Todos os pacientes referiram aumento da dor; três tiveram maior edema e dois piora da astenia; três tiveram infecções de pele e/ou tecido subcutâneo da cabeça e pescoço e um infecção do trato respiratório superior. No 2º grupo (600ug) três pacientes faleceram antes dos 90 dias de protocolo por causas não relacionadas ao tratamento, descritas a seguir: complicações de gastrostomia, rápida progressão tumoral e hemorragia fatal e carcinomatose pulmonar. Neste grupo um paciente não teve EA; cinco tiveram piora da dor; quatro aumento do edema; dois maior astenia; três celulite de face e dois apresentaram sinusite aguda. Como neste grupo observou-se toxicidade classificada como relacionada ao tratamento graus 3 e 4 em mais de um terço dos pacientes decidiu-se escalonar para baixo a dose no 3º grupo para 400ug. No último grupo três pacientes morreram por progressão tumoral rápida, sem completar o protocolo, então incluiu-se três novos indivíduos. Todos tiveram maior dor; três aumento do edema; três piora da astenia; um caso de erisipela; três com infecção pulmonar e um com sinusite aguda; um paciente apresentou linfonodomegalia peri-tumoral após receber a vacina. Concluiu-se que a administração desta vacina neste grupo de pacientes é segura na dose de 400ug e tem como principal toxicidade aumento dos sinais inflamatórios na lesão tumoral e maior ocorrência de infecções locais e respiratórias. / This study goal was to describe and graduate the local and systemic toxicity of intratumoral injections of HSP (Heat Shock Protein) 65 DNA vaccine in advanced-stage head and neck squamous cell carcinoma (SCCHN) patients and to define the highest well tolerated dose for them. This is a prospective, non-randomized, uncontrolled, phase 1 study, using a new vaccine to treat eighteen patients with advanced-stage SCCHN patients without any option for curative treatment, Karnofsky performance status greater than 70% and no organ failure. The patients were divided into 3 groups of 6 patients each one, receiving different vaccine doses, 150ug; 600ug e 1200ug per dose, respectively. They received three injections with a 21 days interval. The patients were rigorously evaluated into their clinical and laboratory aspects looking for adverse events (AE) during 90 days. Toxic effects were monitored according to National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE). In the first group (150ug) one patient died due to an ulcerate lesion extensive bleeding before the protocol end. All the patients referred pain increase; tree had greater edema; two had strong fatigue; tree presented with cutaneous infections of head and neck and one with an acute sinusitis. In the second group (600ug) three patients died before the 90th day protocol, all considered unrelated to treatment, following described: gastrostomy complications, a fatal bleeding after rapid progression of the tumor and pulmonary carcinomatosis. In this group one patient didn\'t have adverse events; five had pain worsening; four had increase of edema; two had greater fatigue; tree had facial cellulitis and two had acute sinusitis. Due to grade 3 and 4 adverse events occurred in more than one third of patients of this group we decided to lower the dose of the third group to 400ug. In the last group three patients also died before protocol completion, all due to cancer progression, so we had to include three more patients in this group. All of them presented greater pain; tree had increasing of edema; tree had fatigue worsening; tree had pulmonary infections; one had acute sinusitis and lymphoadenomegalia besides the tumor lesion. We concluded that intratumoral injections of M.leprae HSP 65 DNA in advanced-stage SCCHN patients is safe at the dose of 400ug/injection and cause tumor flare adverse events and an increase rate of local and respiratory infections.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Cancer vaccines have shown little success in clinic. Dendritic cells (DCs) are of particular interest in cancer vaccination due to their role in cell-mediated immunity. Active targeting of DCs, through PLGA nanoparticles (PLGA-NPs) decorated with ligands for DC-expressed mannose receptor (MR) can enhance internalization, processing and presentation of antigens and subsequent immnuostimulation. In this study we have shown PLGA-NPs decorated with mannan and the synthetic hydrophobized mannan, especially those with covalent attachment, can target DCs leading to increased uptake of nanoparticles and DC maturation. This approach may be used for improved delivery of antigens and adjuvants to DCs and development of more efficient cancer vaccines. Moreover, significant progress in cancer vaccination requires immunomonitoring. Live imaging using a Positron Emission Tomography (PET) probe encapsulated in PLGA-NPs can elucidate dynamics of recruitment and fate of DCs to develop successful vaccines. The PET-nanoprobe prepared by radio-iodinated 5-IDFPdR demonstrated uncontrolled high burst release implying low quality images. / Pharmaceutical Sciences

Thesis (M.Sc.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science in Pharmaceutical Sciences. Title from pdf file main screen (viewed on February 17, 2010). Includes bibliographical references.

No description available.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.

Thesis (Ph. D.)--Ohio State University, 2007. / Full text release at OhioLINK's ETD Center delayed at author's request.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 5 uppsatser.