Fatigue is a common problem among cancer patients, especially those who have received chemotherapy and radiation therapy. Stem cell transplant (SCT) patients are at a particular risk of persistent fatigue as they receive more aggressive therapies. This study examined the prevalence of fatigue after completion of SCT. Further, the level of fatigue symptom distress and its relationship with quality of life (QOL) among long term SCT survivors was examined. The study involved thirty-three patients, 21 males and 12 females, treated with autologous or allogeneic SCT in a comprehensive cancer center in Southwest Florida. Participants' ages ranged from 36 to 70 years, with a mean age of 53 years. All subjects completed the Cancer Related Fatigue Distress Scale and the Functional Assessment of Cancer Therapy-Bone Marrow Transplant questionnaires. All the patients had to be at least six months from transplant. The results of this study showed that fatigue is quite prevalent among SCT survivors. Ninety-three percent of the patients reported some degree of fatigue, and 15% experienced severe fatigue. Patients who received autologous transplant (24%) reported less fatigue symptom distress (mean= 48, SD= 36.62) compared to the allogeneic transplant group (mean= 66.2, SD= 54.49). A strong negative relationship was found between fatigue symptom distress and QOL (r = 0.85, p < 0.0001) suggesting that patients with the greatest fatigue distress report the worst QOL. The time from transplant factor was significantly positively associated with fatigue symptom distress (r= 0.46, p= 0.007) indicating greater distress with the passage of time. A moderate negative relationship was also found between time from transplant and QOL (r= -0.34, p= 0.052) suggesting that QOL was less in some patients as time passed; however this was a weak relationship that did not achieve statistical significance. Although the sample size was small, this study was able to provide a confirmation that fatigue symptom distress and QOL are related to one another. Understanding the relationship between fatigue symptom distress and QOL should encourage interdisciplinary collaboration in planning proper interventions to minimize fatigue. This would improve the outcomes of SCT long term survivors, and would positively impact their overall QOL.

No presenta presenta resumen. / Revisión por pares

Adult bone marrow contains a population of mesenchymal stem cells capable to self-renew and to differentiate into haematopoietic-supportive stroma, osteo, adipo- and chondrocytes. However, the identity of mesenchymal stem cells still remains uncertain. The complex population of their descendants, bone marrow mesenchymal stromal cells (BM MSCs), represents a model to study the principles of differentiation and commitment into mesodermal lineages. The experiments using BM MSCs are often hampered by their low proliferative capacity in vitro. In the present study, we established conditionally immortalized BM MSCs from tetracycline-regulated SV40 Large T-antigen transgenic mice. The identity of the conditionally immortalized BM MSCs was confirmed by marker expression, ability to support haematopoiesis and differentiation potential. The advantages of the conditional immortalization are encompassed in (1) indefinite expansion of cell populations, (2) possibility to perform cellular cloning and (3) prevention from spontaneous differentiation. We demonstrated the heterogeneity of BM MSCs and identified at least 6 types of progenitors within BM MSCs population based on their differentiation potential (“OAC”, “OA”, “OC”, “AC”, “O”, “A”). A hypothetical model of BM MSC hierarchy and the relationships between the progenitors has been proposed. We observed that the Wnt/β-catenin signaling pathway and GSK3 activity could modulate the efficiency of osteo- and adipogenic differentiation pathways, but we didn’t find evidence that the lineage commitment of BM MSCs is determined by Wnt. We elucidated the mechanism of transcriptional regulation of the adipogenic induction of BM MSCs in vitro. Our data revealed the key regulatory role of PPARγ1 during adipogenesis in BM MSCs. Furthermore, we assume that PPARγ1 is a potential trigger of the adipogenic commitment of the BM MSCs progenitors. Finally, the non-adipogenic BM MSCs progenitors were converted into the adipogenic lineage using ectopical expression of the transcription factors C/EBPα, C/EBPβ and C/EBPδ. Our findings provide a novel insight into the molecular mechanisms of BM MSCs lineage commitment.

Lymphatic metastasis in breast cancer (BC) is one of the most important prognostic factors for patient survival. The escaped tumor cells reach distant vital organs and their unopposed expansion in these organs may cause mortality to patient. Tumor cells are transported to lymph node (LN) exclusively by tumor lymphatic vessels (LV). Increased tumor lymphangiogenesis, i.e., the formation of new LV is currently thought to be promoted by soluble factors such as VEGF-C and –D that activate VEGFR-3 expressed in lymphatic endothelial cells (LEC). These factors are secreted by malignant, tumor-infiltrating immune and stromal cells and create a favorable environment for formation of new vessels. However, emerging evidence suggests that tumor lymphangiogenesis is also promoted by Myeloid-derived Lymphatic Endothelial Cell Progenitors (M-LECP). We recently showed that M-LECP are abundant in mouse and human breast tumors and that their density strongly correlates with both lymphatic formation and nodal metastasis. Characterization of M-LECP showed that nearly all these cells express typical markers of the M2-type of macrophages such as CD163, CD204, and CD209. These cells are consider to be strongly immunosuppressive as exemplified by their inhibition of mobilization, activation, and survival of the key defenders against cancer cells, cytotoxic CD8+ T lymphocytes. Here, we compare the in vitro differentiation of M-LECP derived from bone marrow (BM) myeloid precursors primed with CSF-1 followed by secondary stimulants such as LPS, an immunomodulatory ligand for TLR4, and IL-4, IL-13, and IL-10 downstream targets of this receptor that are known to promote M2-macrophage development. Expression of these stimulants was analyzed by qPCR, flow cytometry, and ELISA during M-LECP differentiation. Our study describes the expression and functionality of these Th2 cytokines and their receptors during M-LECP differentiation. We found that each of the Th2 pathways singularly promotes M-LECP differentiation but there is an absent additive effect. We also found that IL-10 but no other Th2 cytokines is upregulated along with its receptor and contributes to the expression of the lymphatic properties similarly to LPS. To our knowledge, the role of IL-10 in development of lymphatic phenotype through differentiation of M-LECP has not been reported previously. Lastly, we show recruitment of M-LECP in a mouse BC model and the co-expression of the Th2 cytokine receptors in these cells. These studies have a potential to identify new regulators of M-LECP production in the bone marrow that could serve as biomarkers and targets for inhibiting tumor lymphatic formation, and by extension, lymph node metastasis.

Both humans and mice display an age-related decline in immunity. Reduced generation of mature B cells may be a contributing factor due to reduced entry of mature B cells with novel B cell receptors and specificity for pathogens into the mature B cell pool. In aged mice the numbers of B cell precursors within the bone marrow are diminished; there is a severe reduction in numbers of pre-B cells and an increase in numbers of re-circulated mature B cells. Other defects in developing B cells include reduced expression of rag1 and rag2 when measured in total bone marrow precursor populations. In the pro-B cell stage of development rag expression is essential to the process of V(D)J recombination and the generation of pre-B cells. It was not known prior to this work if rag levels were lower in pro-B cells. In Chapter 2 I show that rag2 expression is reduced in pro-B cells of aged mice. The reduction in rag2 expression is correlated with a loss of V(D)J recombinase activity in pro-B cells and reduced numbers of pre-B cells. This suggests that in aged mice the reduction in rag2 expression is sufficient to result in reduced V(D)J recombinase activity and reduced generation of pre-B cells, thus contributing to fewer pre-B cells in aged mice. Furthermore, I have shown that the loss of rag2expression and recombinase activity in pro-B cells are the result of age-associated defects in the bone marrow-microenvironment as opposed to cell-intrinsic defects in developing precursors. In Chapter 3 of this thesis I examine genetic influences on age-related defects in murine B cell development and correlations between bone marrow B cell subsets and peripheral T cell subsets. It was known that longevity and age-related defects in T cell subsets are influenced by genetic differences between strains of inbred mice. The impact of genetic polymorphisms on age-related defects in B cell development had not been previously assessed. Nor was it known if these defects were correlated with age-related changes in peripheral T cell subsets. Here I present evidence that B cell subsets in the bone marrow are influenced by genetic polymorphisms between mice strains. Genetic polymorphisms on Chromosomes 15 and 19 were found to influence the frequency of re-circulated and pre-B cells in the bone marrow of aged mice. Frequencies of bone marrow B cell subsets were compared with peripheral T cell subsets. Interestingly, an association between the frequency of pre-B cells was not observed with either re-circulated B cells in the bone marrow nor peripheral T cell subsets. However the frequency of pre-B cells was inversely correlated with the frequency of B220intIgM+cells, a subset that was found to correlate with more advanced age-related T cell defects. In addition, frequencies of re-circulated B cells in the bone marrow were found to be associated with less advanced age-related defects in peripheral T cell subsets. These observations indicate that defects in B cell development, including reduced rag2 expression and V(D)J recombinase activity, are the result of changes in the aged murine bone marrow microenvironment. In addition, a genetic polymorphism located on Chromosome 19 influences the frequency of pre-B cells in aged mice. Furthermore the frequencies of B cell precursors in aged mice are not correlated with peripheral T cell subsets, but are correlated with frequencies of B220intIgM+ cells in the bone marrow. These observations advance our understanding of age-related defects in murine B cell development and may lead to identification of genes that influence B cell development in aged mice and humans as well as to help devise therapeutics aimed at restoring humoral immunity in aged individuals.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第17978号 / 医博第3842号 / 新制||医||1001(附属図書館) / 80822 / 京都大学大学院医学研究科医学専攻 / (主査)教授 長澤 丘司, 教授 河本 宏, 教授 江藤 浩之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医科学) / 甲第18906号 / 医科博第62号 / 新制||医科||4(附属図書館) / 31857 / 京都大学大学院医学研究科医科学専攻 / (主査)教授 山下 潤, 教授 野田 亮, 教授 髙折 晃史 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19398号 / 医博第4049号 / 新制||医||1012(附属図書館) / 32423 / 京都大学大学院医学研究科医学専攻 / (主査)教授 戸口田 淳也, 教授 妻木 範行, 教授 井上 治久 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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