Ko Kin Ming Jeffery. / Thesis submitted in: July 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 99-111). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Table of Contents --- p.ix / List of Tables --- p.xiii / List of Figures --- p.xv / List of Abbreviations --- p.xvi / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Malignant Lymphoma --- p.1 / Chapter 1.2 --- Non-Hodgkin's Lymphoma --- p.1 / Chapter 1.3 --- Diffuse Large B-cell Lymphoma --- p.2 / Chapter 1.3.1 --- General Features of Diffuse Large B-cell Lymphoma --- p.2 / Chapter 1.3.2 --- Morphologic variants of Diffuse Large B-cell Lymphoma --- p.4 / Chapter 1.3.2.1 --- Centroblastic vairant --- p.5 / Chapter 1.3.2.2 --- Immunoblastic variant --- p.5 / Chapter 1.3.2.3 --- Anaplastic variant --- p.6 / Chapter 1.3.3 --- Immunophenotype of Diffuse Large B-cell Lymphoma --- p.6 / Chapter 1.3.3.1 --- Lineage-associated antigens --- p.6 / Chapter 1.3.3.1.1 --- B-cell lineage antigens --- p.6 / Chapter 1.3.3.1.2 --- T-cell lineage antigens --- p.7 / Chapter 1.3.3.2 --- Antigen involved in regulation of cell proliferation and apoptosis --- p.8 / Chapter 1.3.3.2.1 --- Proliferation markers --- p.8 / Chapter 1.3.3.2.2 --- Cell cycle regulators --- p.8 / Chapter 1.3.3.2.3 --- Protein controlling apoptosis --- p.10 / Chapter 1.3.4 --- Subtypes of Diffuse Large B-cell Lymphoma --- p.10 / Chapter 1.3.4.1 --- Classification method of DLBCL subtypes --- p.11 / Chapter 1.3.4.1.1 --- DNA microarray --- p.11 / Chapter 1.3.4.1.2 --- Immunohistochemistry pattern --- p.14 / Chapter 1.3.4.1.2.1 --- CD10 --- p.16 / Chapter 1.3.4.1.2.2 --- Bcl-6 --- p.16 / Chapter 1.3.4.1.2.3 --- CD138 --- p.17 / Chapter 1.3.4.1.2.4 --- MUM1/IRF4 --- p.17 / Chapter 1.3.4.2 --- Prognosis of 、DLBCL subtypes --- p.19 / Chapter 1.4 --- Interleukin 10 --- p.22 / Chapter 1.4.1 --- The IL-10 gene --- p.23 / Chapter 1.4.2 --- IL-10 promoter --- p.23 / Chapter 1.5 --- IL-10 receptor --- p.24 / Chapter 1.6 --- Cellular Signaling Pathways Regulated by IL-10 --- p.25 / Chapter 1.6.1 --- Jak/Stat Pathway --- p.25 / Chapter 1.6.2 --- Inhibition of NF B pathway --- p.26 / Chapter 1.7 --- Function of IL-10 --- p.27 / Chapter 1.7.1 --- Effects of IL-10 on immune cells in vitro --- p.27 / Chapter 1.7.2 --- Effects of IL-10 on B-cells --- p.28 / Chapter 1.8 --- IL-10 and IL-10 receptor in malignant diseases --- p.29 / Chapter 1.8.1 --- Melanoma --- p.29 / Chapter 1.8.2 --- Carcinoma --- p.30 / Chapter 1.8.3 --- Lymphoma --- p.30 / Chapter 1.9 --- Single Nucleotide Polymorphism (SNP) --- p.33 / Chapter 1.9.1 --- SNPs in cancer research --- p.34 / Chapter 1.9.1.1 --- Susceptibility to cancer and SNPs --- p.35 / Chapter 1.9.1.2 --- Outcome and SNPs --- p.35 / Chapter 1.10 --- SNP in the IL-10 promoter --- p.36 / Chapter 1.11 --- IL-10 promoter SNP in DLBCL --- p.37 / Chapter Chapter 2: --- Aims of Study --- p.39 / Chapter Chapter 3: --- Materials and Methods --- p.41 / Chapter 3.1 --- Sample Recruitment --- p.41 / Chapter 3.2 --- DNA preparation for Single Nucleotide Polymorphism (SNP) analysis --- p.41 / Chapter 3.2.1 --- Isolation of Peripheral Blood Mononuclear Cell (PBMC) from buffy coat from blood of normal control group --- p.41 / Chapter 3.2.2 --- Preparation for NHL and DLBCL samples from paraffin-embedded sections for DNA extraction --- p.42 / Chapter 3.2.3 --- DNA extraction for SNP analysis --- p.42 / Chapter 3.3 --- SNP analysis by Restriction Fragment Length Polymorphism (RFLP) --- p.43 / Chapter 3.3.1 --- Amplification of target site by PCR --- p.43 / Chapter 3.3.2 --- SNP analysis --- p.45 / Chapter 3.4 --- Determination of haplotypic frequency --- p.50 / Chapter 3.5 --- Classification of DLBCL by immunohistochemistry --- p.50 / Chapter 3.5.1 --- Staining pattern of CD10 --- p.53 / Chapter 3.5.2 --- Staining pattern of Bcl-6 --- p.54 / Chapter 3.5.3 --- Staining pattern of CD138 --- p.55 / Chapter 3.5.4 --- Staining pattern of MUM1/IRF4 --- p.56 / Chapter 3.6 --- Statistical Analysis --- p.57 / Chapter Chapter 4: --- Results --- p.58 / Chapter 4.1 --- SNPs of IL-10 promoter in normal controls --- p.58 / Chapter 4.1.1 --- Allelic Frequencies and genotype distributions --- p.58 / Chapter 4.1.2 --- Haplotypic Frequencies of normal controls --- p.58 / Chapter 4.2 --- SNP of the IL-10 promoter in non-Hodgkin's lymphomas --- p.59 / Chapter 4.2.1 --- Allelic frequencies and genotype distributions --- p.59 / Chapter 4.2.2 --- Haplolypic frequencies --- p.61 / Chapter 4.4 --- SNPs of the IL-10 promoter in DLBCL --- p.62 / Chapter 4.4.1 --- Allelic frequencies and genotype distributions --- p.62 / Chapter 4.4.2 --- Haplotypic frequencies --- p.64 / Chapter 4.5 --- SNP of the IL-10 promoter in different subtypes of DLBCL --- p.65 / Chapter 4.5.1 --- Classification of DLBCL by immunohistochemistry --- p.65 / Chapter 4.5.2 --- SNP of the IL-10 promoter in Germinal Center DLBCL (GC-DLBCL) --- p.67 / Chapter 4.5.2.1 --- Allelic frequencies and genotype distributions --- p.67 / Chapter 4.5.1.2 --- Haplotypic frequencies --- p.69 / Chapter 4.5.2 --- SNP of the IL-10 promoter in Activated Germinal Center DLBCL (AGC-DLBCL) --- p.70 / Chapter 4.5.2.1 --- Allelic frequencies and genotype distributions --- p.70 / Chapter 4.5.2.2 --- Haplotypic frequencies --- p.72 / Chapter 4.5.3 --- SNP of the IL-10 promoter in Activated non-Germinal Center DLBCL (ANGC-DLBCL) --- p.73 / Chapter 4.5.3.1 --- Allelic frequencies and genotype distributions --- p.73 / Chapter 4.5.3.2 --- Haplotypic frequencies --- p.75 / Chapter 4.5.4 --- SNP of the IL-10 promoter in Unclassified DLBCL (UC-DLBCL). --- p.76 / Chapter 4.5.4.1 --- Allelic frequencies and genotype distributions --- p.76 / Chapter 4.5.4.2 --- Haplotypic frequencies --- p.78 / Chapter 4.6 --- Summary of SNP of the IL-10 promoter in DLBCL subtypes --- p.79 / Chapter 4.7 --- Overall survival analysis --- p.80 / Chapter 4.7.1 --- Clinical data of DLBCL --- p.80 / Chapter 4.7.2 --- Cox Proportional Hazards Regression Analysis in DLBCL --- p.81 / Chapter Chapter 5: --- Discussion --- p.88 / Chapter 5.1 --- SNP for low IL-10 production in Hong Kong population --- p.88 / Chapter 5.2 --- NHL in low IL-10 production population --- p.90 / Chapter 5.2.1 --- The relationship between IL-10 and NHL --- p.90 / Chapter 5.2.2 --- Allelic frequencies and haplotype of the IL-10 promoter in NHL --- p.90 / Chapter 5.3 --- Classification of DLBCL --- p.91 / Chapter 5.3.1 --- Current prognostic analysis --- p.91 / Chapter 5.3.2 --- DLBCL subtypes distribution in Hong Kong is different from Caucasian --- p.92 / Chapter 5.4 --- IL-10 and DLBCL --- p.93 / Chapter 5.5 --- SNP of IL-10 promoter in DLBCL subtypes --- p.94 / Chapter 5.5.1 --- Allelic frequencies and haplotype of DLBCL subtypes --- p.94 / Chapter 5.5.2 --- Rare haplotypes were discovered in DLBCL --- p.94 / Chapter 5.6 --- Overall survival Analysis --- p.95 / Chapter 5.6.1 --- Univariate Cox Proportional Hazards Regression Analysis --- p.95 / Chapter 5.6.2 --- Bivariate Cox Proportional Hazards Regression Analysis --- p.96 / Chapter Chapter 6: --- Conclusion --- p.97 / References --- p.99

Ces travaux de thèse présentent une approche de pharmaco-épidémiologie explorant l'exposition médicamenteuse chez des patients atteints de lymphome tout au long de leur parcours de soin. L'utilisation des données du Système National d'Information Inter-régimes de l'Assurance Maladie (SNIIRAM) nous a permis de quantifier cette exposition médicamenteuse et de déterminer les facteurs associés dans le contexte de la vie réelle. Les travaux de recherche réalisés au cours de cette thèse répondaient à 3 objectifs. Dans un premier objectif, nous avons souhaité valider l'utilisation du SNIIRAM à des fins de recherche pour améliorer la robustesse des futures études menées sur le lymphome. Après avoir défini plusieurs algorithmes d'identification des cas incidents de lymphome, nous avons testé leur validité par confrontation aux données cliniques exhaustives du Registre des cancers du Tarn. Les paramètres de performance obtenus permettent de considérer les données disponibles dans le SNIIRAM comme un outil puissant pour mener des études pharmaco-épidémiologiques ou médico-économiques sur le Lymphome. Le second objectif était d'explorer l'existence d'une surconsommation de médicaments psychotropes au cours de la phase active de traitement par rapport à une population témoin, sous l'hypothèse d'une initiation accrue de ces médicaments pour pallier les complications psychologiques associées à la prise en charge du Lymphome. Par ailleurs, la chronicisation fréquemment observée de ce type de consommation peut conduire à terme à des complications potentiellement évitables. Pour répondre à cet objectif, nous avons réalisé une étude selon une approche " new-user design ", à partir d'une cohorte de patients incidents de lymphome identifiés dans les données du SNIIRAM à l'échelon régional. Nous avons observé un taux d'initiation supérieur au taux observé en population générale ou chez des patients atteints d'autres pathologies mettant en jeu le pronostic vital (infarctus du myocarde). En fonction de la classe thérapeutique étudiée, 20 à plus de 50% des patients restaient exposés de façon inappropriée (au-delà des recommandations) à ces médicaments. Le troisième objectif a donc été d'explorer les déterminants associés à une utilisation prolongée de médicaments psychotropes dans la phase de l'après-cancer à partir des données de l'Echantillon généraliste des bénéficiaires (EGB). [...] / This thesis presents a pharmacoepidemiology approach to describe drug utilization in lymphomas during their whole care pathway. The use of the French claims database (Système National d'Informations Inter-Régimes de l'Assurance Maladie (SNIIRAM)) allows to exhaustively quantify this drug utilization in real life conditions. This thesis consists of three mains objectives. First, we aimed to develop validated algorithms for the identification of incident cases of lymphoma. For the validation, we used data from a regional Cancer Registry as the gold standard. The purpose of this validation study was to enhance validity of future studies conducted on lymphomas in the SNIIRAM database. The results of this study associated to strengths of this database demonstrate that this approach is of great interest to conduct pharmacoepidemiological or medico-economic studies in lymphomas. Second, we aimed to estimate the incidence of use of psychotropic drugs during the active treatment phase of lymphoma in comparison with control groups. Indeed, the increased probability of developing anxio-depressive disorders after diagnosis could lead to an increased initiation of psychotropic drugs and a potential inappropriate chronic use of these drugs after initiation. Such inappropriate chronic use can unnecessarily expose patients to adverse event. For this aim, we conducted a new-user cohort study using data from the SNIIRAM database. The results of this study revealed that the initiation rate of these drugs is higher than in the general population or for life-threatening diseases such as myocardial infarction. Moreover, we observed an inappropriate prolonged use for a significant fraction of patients (20% to more than 50% according to therapeutic class). On the basis of these findings, the third objective was to identify factors associated with prolonged use of these drugs during survivorship. This study was conducted using data from the General Sample of Beneficiaries (EGB). [...]

Thesis (Ph. D.)--Ohio State University, 2007. / Title from first page of PDF file. Includes bibliographical references (p. 176-216).

El limfoma de cèl•lules del mantell (LCM) és una neoplàsia heterogènia en relació a les característiques clíniques, biològiques i moleculars que presenten els pacients. La morfologia, l’immunofenotip i el genotip no són suficients per explicar aquesta gran diversitat. Encara que es considera una neoplàsia agressiva alguns pacients segueixen un curs clínic indolent sense la necessitat de tractament en el moment del diagnòstic i presenten una supervivència llarga. Els resultats d’aquesta tesi demostren que els LCM amb elevada càrrega mutacional en els gens de les immunoglobulines (IGHV)(<97% d’identitat amb la línia germinal) utilitzen preferencialment certes famílies de gens IGHV, suggerint la presència de processos de selecció antigènica com a mecanisme de patogènesi d’aquesta neoplàsia i donant importància a la caracterització del receptor de les cèl•lules B (BCR). Els LCM amb gens IGHV mutats (M-LCM) presenten una menor complexitat genòmica, són preferencialment negatius per a l’expressió del factor de transcripció SOX11 i presenten afectació ganglionar amb més freqüència que els casos sense mutacions (U-LCM). Per altra banda, el perfil d’expressió gènica entre casos amb i sense mutacions en els gens IGHV, també mostra expressió diferencial de programes cel•lulars relacionats amb poblacions de cèl•lules B normals incidint amb la ontogènia d’ambdós grups. El perfil d’expressió de microRNAs també permet diferenciar entre dos grups de pacients amb diferent supervivència, reforçant la idea de la presència de dos subgrups que pertanyen a una mateixa entitat. La identificació i la correcte identificació d’aquests pacients de limfoma de cèl•lules del mantell són rellevants per al posterior maneig i tractament dels mateixos. / Mantle cell lymphoma is a malignant and heterogeneous disease regarding the clinical, biological and molecular characteristics presented by patients, all of them with the translocation t(11;14) and CCND1 overexpression. Although is considered an aggressive neoplasm some patients follow and indolent clinical course without treatment at the time of diagnosis and long survival. The results of this studies demonstrate that LCM with high mutational load in the immunoglobulins genes (IGHV) (<97% of identity with the germ line) use preferentially specific IGHV gene families, suggesting the presence of antigenic selection process as a mechanism of pathogenesis of this neoplasm and giving importance to the characterization of the B cell receptor. The LCM cases with IGHV mutated genes (M-LCM) present less genomic complexity are preferentially negative for the expression of the transcriptor factor SOX11 and also present lymphadenopathies more frequently compared with the IGHV unmutated cases (U-LCM). Furthermore, the gene expression profile between cases with and without IGHV mutations also shows differential expression of cellular programs related to normal B cell populations maybe related with the ontogeny for both groups. The expression profile of microRNAs also differentiates between two groups of patients with different survival and other molecular characteristics, reinforcing the idea of the presence of two subgroups that belong to the same entity. The correct identification and classification of these patients are relevant for subsequent handling and treatment of the patients.

Los resultados de esta tesis doctoral han permitido profundizar en el reconocimiento y clasificación del linfoma de células del manto (MCL), basándonos en dos subtipos poco habituales. Por un lado, hemos estudiado las muestras de los pacientes de MCL que no presentan la traslocación principal t(11;14), característica que ha sido objeto de discusión al pensar que el diagnóstico de MCL podía no ser el correcto. Por otro lado, hemos estudiado los MCL con presentación clínica indolente, larga supervivencia, y que no requieren tratamiento durante largos períodos de tiempo, particularidad llamativa en los pacientes de MCL. En las dos primeras publicaciones de esta tesis hemos profundizado en el estudio de los MCL ciclina D1 negativos. Hemos demostrado que el factor de transcripción SOX11, además de sobreexpresarse en MCL ciclina D1 positivos, es un biomarcador útil para identificar los MCL ciclina D1 negativos. Además, caracterizamos genéticamente estos casos y mostramos que el 55% presenta traslocaciones de la CCND2 predominantemente con cadenas ligeras de los genes de las inmunoglobulinas. El perfil de alteraciones genómicas estudiado con arrays de alta densidad ha demostrado que las alteraciones genéticas secundarias entre los MCL ciclina D1 negativos y positivos son muy similares. Los casos de MCL ciclina D1 negativos tienen características clínicas y patológicas similares a los ciclina D1 positivos. Las translocaciones de CCND2 junto con la expresión de SOX11 son herramientas de diagnóstico muy útiles en la identificación de los MCL ciclina D1 negativos para que éstos puedan beneficiarse de los mismos protocolos terapéuticos usados en los MCL convencionales. En el tercer trabajo de esta tesis, nos hemos centrado en el reconocimiento del grupo de pacientes de MCL que no necesita tratamiento durante largos periodos de tiempo, idea que está modificando la manera de entender el MCL, ya que en general está considerado un linfoma agresivo que requiere un tratamiento inmediato. En este trabajo nos basamos en el estudio de Fernandez et al.1 en que se comparó un grupo de pacientes de MCL con comportamiento clínico indolente con un grupo de pacientes con MCL convencional (agresivo). Ambos grupos de muestras tenían un perfil de expresión global similar, sugiriendo que pertenecen a la misma enfermedad, pero también diferían en la expresión de una pequeña firma de 13 genes que estaba altamente expresada en los casos convencionales de MCL pero era negativa en los tumores de los pacientes con comportamiento indolente. Por lo que en el presente estudio, diseñamos y validamos un ensayo de PCR cuantitativa, seleccionando tres genes de la firma de perfil de expresión génica: SOX11, HDGFRP3 y DBN1 que permiten clasificar los casos en uno de estos dos grupos. El subgrupo con baja expresión de los tres genes incluye principalmente los pacientes con clínica indolente, afectación no-ganglionar, genes de las inmunoglobulinas hipermutados, y una supervivencia significativamente superior en comparación a los casos con expresión alta. Además, los MCL leucémicos sin afectación ganglionar y con alta expresión de los tres genes tienen una supervivencia significativamente inferior y un comportamiento clínico más agresivo que los pacientes con firma de expresión baja sin afectación ganglionar. En cuanto a las alteraciones genéticas, tanto los MCL con alta o baja expresión de los tres genes tenían una supervivencia significativamente inferior si presentan alteraciones en 17p. Por tanto, concluimos que la evaluación de la firma de los tres genes junto con el estudio de 17p/TP53 en muestras leucémicas puede ayudar a identificar un subgrupo particular de MCL y determinar la evolución clínica de estos los pacientes. Por tanto, estos estudios pueden proporcionar una evaluación más precisa del MCL y una ayuda en la decisión de tratamiento más adecuado para cada paciente. / In the first publication of this thesis we have studied the cyclin D1-negative mantle cell lymphomas (MCL). These cases are not well characterized due to the difficulties in their recognition. Overexpression of the transcription factor SOX11 has been observed in conventional MCL. In this study we demonstrated that SOX11 was highly expressed in cyclin D1-negative MCL, thus SOX11 expression is a highly specific marker for both cyclin D1-positive and negative MCL. In the second study we investigated 40 cyclin D1-negative MCL. Chromosomal rearrangements of CCND2 were detected in 55% of the cases and frequently with light chain immunoglobulin (IG) genes as partners. The genomic profile of the cyclin D1-negative cases analyzed by high resolution arrays were similar to the cyclin D1-positive MCL. This characterization of a large series of cyclin D1-negative MCL indicates that these tumors are clinically and biologically similar to the cyclin D1-positive MCL and provides a basis for the proper identification and clinical management of these patients. In the third publication, we studied the clinical and biological features of mantle cell lymphoma with a more indolent disease and long survival. The expression of 3 genes (SOX11, HDGFRP3, DBN1) selected from a gene expression signature distinguishing conventional an indolent MCL was studied in 68 leukemic MCL by quantitative PCR. An unsupervised analysis segregated two groups of MCL based on the expression levels of these genes. The tumors with low expression presented mainly with a non-nodal disease, had more frequently mutated IG, fewer genomic alterations, remained untreated more frequently and longer time, and had a better outcome than the tumors with high expression. The presence of 17p/TP53 alterations had an adverse effect on the outcome of both subgroups of MCL. The detection of the 3-gene signature may help to identify this particular subtype of MCL.

published_or_final_version / Pathology / Master / Master of Medical Sciences

Cell proliferation is a process that is strictly regulated by a large number of proteins. An alteration in one of the encoding genes inserts an error into the regulative protein, which may result in uncontrolled cell growth and eventually tumor formation. Lymphoma is a cancer type originating in the lymphocytes, which are part of the body’s immune defence. In the present thesis, Znfn1a1, Notch1 and Bcl11b were studied; all involved in the differentiation of T lymphocytes. The three genes are located in chromosomal regions that have previously shown frequent loss of heterozygosity in tumor DNA. Ikaros is a protein involved in the early differentiation of T lymphocytes. In this thesis, mutation analysis of the Znfn1a1 gene in chemically induced murine lymphomas revealed point mutations and homozygous deletions in 13 % of the tumors. All of the detected deletions lead to amino acid substitutions or abrogation of the functional domains in the Ikaros protein. Our results support the role of Ikaros as a potential tumor suppressor in a subset of tumors. Notch1 is a protein involved in many differentiation processes in the body. In lymphocytes, Notch1 drives the differentiation towards a T-cell fate and activating alterations in the Notch1 gene have been suggested to be involved in T-cell lymphoma. We identified activating mutations in Notch1 in 39 % of the chemically induced murine lymphomas, supporting the involvement of activating Notch1 mutations in the development of T-cell lymphoma. Bcl11b has been suggested to be involved in the early T-cell specification, and mutations in the Bcl11b gene has been identified in T-cell lymphoma. In this thesis, point mutations and deletions were detected in the DNA-binding zinc finger regions of Bcl11b in 15 % of the chemically induced lymphomas in C57Bl/6×C3H/HeJ F1 mice. A mutational hotspot was identified, where four of the tumors carried the same mutation. Three of the identified alterations, including the hotspot mutation in Bcl11b, increased cell proliferation when introduced in a cell without endogenous Bcl11b, whereas cell proliferation was suppressed by wild-type Bcl11b in the same cell line. Mutations in Bcl11b may therefore be an important contributing factor to lymphomagenesis in a subset of tumors. A germ line point mutation was identified in BCL11B in one of 33 human B-cell lymphoma patients. Expression of BCL11B in infiltrating T cells was significantly lower in aggressive compared to indolent lymphomas, suggesting that the infiltrating T cells may affect the B-cell lymphomas.

Thesis (Ph. D.)--University OF MICHIGAN. / Also issued in print.

Thesis (Ph. D.)--University OF MICHIGAN. / eContent provider-neutral record in process. Description based on print version record.

Thesis (Ph. D.)--University OF MICHIGAN.