ABSTRACT THE EFFECTS OF HIV DISEASE AND LIFESTYLE FACTORS ON CELLULAR AGING IN TRANSGENDER WOMEN by Scott Stephen Sohn Background: Telomeres are short tandem repeats of nucleotides at the ends of chromosomes. These specialized structures serve as caps on the end of the chromosomes, which protect DNA integrity. Telomeres get shorter each time a cell replicates, but the DNA remains intact as long as the telomere caps are a sufficient length. In time, telomeres become too short to protect DNA, which leads to cellular death. Previous research has shown that disease and negative lifestyle factors play a role in accelerated telomere attrition throughout the cellular life cycle. Objective: The purpose of this study was to determine if HIV infection and lifestyle factors in a transgender population living in Atlanta Georgia are associated with telomere length reduction. Participants/setting: This study is a secondary analysis of data provided by a Georgia State University study entitled “Telomere Length, Environmental Stressors and Health Related Outcomes among Transgender Women”. The study included 92 transgender women from Atlanta, Georgia with 49 reporting HIV infection. Two sources of data were collected, survey responses collected during face to face interviews and a saliva sample for DNA analysis. Statistical analysis: Frequency statistics were used to describe the sample population. A Mann Whitney U was used to evaluate telomere length using the T/S ratio by HIV status, by physical activity level (healthy active or low active) and by fruit and vegetable intake category (Don’t eat, 1-2 servings/day, 3-4 servings/day vs. >5 servings/day) in the total Population. Multiple regression analysis was used to examine the association between independent variables (activity level, body mass index, fruit and vegetable intake, hormone use, race, HIV status and age) and telomere length. Results: The majority of the population was Black (84%) with a median age of 33 years (range, 18 to 65 years). No significant association was observed between HIV infection and T/S ratio. The vast majority of the population reported low activity level and only 9% reported consuming >5 servings of fruits and vegetables daily. No significant association was found between fruit and vegetable intake or physical activity level and T/S ratio in this population. Conclusion: HIV infection, Fruit and vegetable intake, and physical activity were not found to impact telomere length in an urban population of transgender women. Future research is needed to further understand the mechanisms that impact telomere length throughout the cellular life cycle within the transgender population.

Telomere, the ends of chromosomes, consists of simple hexameric repeats. In human, TTAGGG repeat is found at the ends of all chromosomes. Telomeres progressively shorten with age in somatic cells, because the insufficient telomerase activity fails to compensate the progressive telomeric erosion. Thus, the reduction of telomeric length in senescent cells is believed to result from active cell division that erodes chromosomal termini. The telomere length supposed to have been an evaluation tool for aging in human. Schizophrenia is a thought, perception disorder, generally regarded as an illness with onset in late adolescence or early adult life with a prevalence rate of 1%. Although the long-term course of schizophrenia shows great heterogeneity among patients, a significant number of patients experience very poor outcomes, shows severe cognitive impairment that is suggestive of a progressive neurodegenerative disorder. The aim of this study is to explore the relationship between neurodegenerative process and telomere length in schizophrenic patients. The latency of P300 event-related potentials is prolonged in disorders associated with neural damage and degeneration and also becomes prolonged in the course of aging process. This study is separated as two parts: first part, using the event-related potentials P300 latency as a tool to evaluate the cognitive dysfunction and aging process in schizophrenics. One hundred and fifty three long-term hospitalization chronic schizophrenics were recruited as the experimental group of this research, including 44 male and 109 female patients with mean age of 38.4 years. These patients were divided into 2 groups according to the different responses to treatment, global assessment functional scale (GAF): 91 with good response to treatment; 62 with poor response to treatment. The normal control group included 101 normal people, male 37 and female 76, with mean age of 38.1 years. The event-related potentials was elicited by auditory oddball paradigm. The P300 latency prolonged in these two schizophrenic patients. The longest P300 latency was found in the poor response schizophrenic group. The shortest P300 latency was found in the normal group. Linear regression coefficients were computed to determine the slope of component P300 latency on age and other factors. The slope of P300 latency on age in normal control group is 1.2 ms/y. In schizophrenic groups, not only the age, but the GAF as the most contributing factors in the neurodegenerative process. Second part, the subjects were selected from part one, 48 chronic schizophrenia whose mean age was 37.9 years and 48 age-, handedness-, and gender-matched normal control subjects. The schizophrenic patients were divided into 2 groups according to the different responses to treatment: 34 with good response to treatment; 14 with poor response to treatment. The telomere length, measured by assay of terminal restriction fragments (TRFs), was determined in HinfI-digested DNA by Southern blot analysis using a (TTAGGG)4 probe. The shortest TRF length was found in the poor response schizophrenic group. There was no difference between the good response schizophrenic group and normal control group. TRF length in peripheral leukocytes obtained from normal control group decreased by approximately 89bP per year. In schizophrenic groups, the TRF length was found that not the age, but the age onset and GAF as the most contributing factors in linear regression model. This study shows that the poor response schizophrenic patients have the most rapid neurodegenerative process in P300 latency and TRF length evaluation. It implicates the homogeneous group, and can be considered as the kraepelinian schizophrenics, very poor outcome group.

Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy. Most ALL cases originate from immature B-cells (BCP-ALL) and are characterized by reoccurring structural genetic aberrations. These aberrations hold information of the pathogenesis of ALL and are used for risk stratification in treatment. Despite increased knowledge of genetic aberrations in pediatric T-cell ALL (T-ALL), no reliable molecular genetic markers exist for identifying patients with higher risk of relapse. The lack of molecular prognostic markers is also evident in patients with relapsed ALL. During the last decades, aberrant epigenetic mechanisms including DNA methylation have emerged as important components in cancer development. Telomere maintenance is another important factor in malignant transformation and is crucial for long-term cell survival. Like DNA methylation, telomere length maintenance has also been implicated to reflect outcomes for patients with leukemia. In this thesis, the prognostic relevance of DNA methylation and telomere length was investigated in pediatric ALL at diagnosis and relapse. The telomere length (TL) was significantly shorter in diagnostic ALL samples compared to normal bone marrow samples collected at cessation of therapy, reflecting the proliferation associated telomere length shortening. Prognostic relevance of TL was shown in low-risk BCP-ALL patients where longer telomeres at diagnosis were associated with higher risk of relapse. Genome-wide methylation characterization by arrays in diagnostic T-ALL samples identified two distinct methylation subgroups denoted CIMP+ (CpG Island Methylator Phenotype high) and CIMP- (low). CIMP- T-ALL patients had significantly worse outcome compared to CIMP+ cases. These results were confirmed in a Nordic cohort treated according to the current NOPHO-ALL2008 protocol.  By combining minimal residual disease (MRD) status at treatment day 29 and CIMP status at diagnosis we could further separate T-ALL patients into risk groups. Likewise, the CIMP profile could separate relapsed BCP-ALL patients into risk groups, where the CIMP- cases had a significantly worse outcome compared to CIMP+ cases.  From these data we conclude that DNA methylation subgrouping is a promising prognostic marker in T-ALL, as well as in relapsed BCP-ALL two groups where reliable prognostic markers are currently missing. By elucidating the biology behind the different CIMP profiles, the pathogenesis of ALL will be further understood and may contribute to new treatment strategies.

Telomeres are located at the ends of all eukaryotic chromosomes and provide the stability of chromosomes. They consist of simple tandem hexametric repeats and play an important part in cell longevity. In human lymphocytes, telomeres shorten progressively with age. Mental retardation (MR) is a disorder with intelligence quotient below average (IQ < 70) and impairment in adaptive skills. IQ by Weschsler Adult Intelligence Scales revised (WAIS-R) appears to peak in the of 30-34 and thereafter decline gradually. Life expectancy is defined as the number of years remaining to be lived. The overall increase in life expectancy indicates an improvement in longevity. The life expectancy of MR patients is shorter than that of the general population. The purpose of this study is to predict the relationship between telomere length and IQ in normal control as well as to analyze the differences among the average telomere length for the control and subgroups of MR cases. Fifty-nine patients who met the fourth edition of Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria for mental retardation were included in this study. According to the degree of intellectual impairment, MR patients were divided into 4 types: mild, moderate, severe, and profound. Fifty-two female nursing students aged 19-21 were recruited as normal controls. DNA was isolated from their lymphocytes. Telomere length was analyzed by Southern blot hybridization. The length was calculated by the Photo CaptMw Version 99.03 software. Correlation between the telomere length and IQ in normal control was performed by the Pearson product-moment correlation. One-way ANOVA was used to test if any differences existed among the normal, mild, moderate, severe, and profound MR. Analyses displayed that there were no correlations between telomere length and IQ including PIQ(r=-0.001; p=0.922), VIQ(r=-0.033; p=0.817), TIQ(r=-0.026, p=0.857), and no difference existed among the normal and subgroups of MR cases. Results obtained from this study indicated that life expectancy of MR patients may approximate to that of the general population if live in the well environment.

Objective: Child maltreatment (CM) is a widespread problem associated with poor mental and physical health outcomes. The underlying mechanisms of this link are not always well understood, however certain biological changes observed in maltreated individuals may play a role in connecting experience and outcome. This review specifically focuses on two markers of biological embedding, DNA methylation (DNAm) and telomere length (TL) in maltreated children and youth. As biomarker changes are not uniform among maltreated children, we additionally discuss biological and environmental resilience factors that may contribute to variability. Methods: We conducted a systematic review of Medline, Embase and PsycInfo databases for studies examining DNAm and/or TL in maltreated children and youth. Methodological quality of the included articles was assessed using the Scottish Intercollegiate Guidelines Network (SIGN) checklists for cohort studies and randomized control trials. Data extraction focused on various factors including population and CM (type, chronicity, severity, and duration) characteristics. Results: The initial search returned 1,688 non-duplicate results, with 417 full text articles reviewed. Twenty-six articles from 16 studies were ultimately included of which 8 examined telomere length and 18 examined DNA methylation. Conclusions: While some heterogeneity of findings was found, evidence supports differential changes in both biomarkers associated with CM. This review enhances understanding of the constellation of biological changes related to CM and consideration of the important role of resilience factors in mitigating risk. Elucidating these factors may highlight targets for future study and intervention development. / Thesis / Master of Science (MSc) / Child maltreatment is a serious problem linked to poor mental and physical health outcomes. The mechanisms of these links are not always clear, however biological changes observed in some maltreated individuals may play a role. Here, we systematically review literature related to two biomarkers of interest in maltreated children, telomere length and DNA methylation. Findings are varied; however, overall, they support an association between child maltreatment and changes in both biomarkers. We additionally discuss factors that may confer resilience related to these changes to highlight potential targets for future study and interventions.

Telomeres form protective caps at the ends of linear chromosomes. They consist of repetitive DNA nucleotides and associated proteins of the shelterin complex. In vitro telomeres become shorter during cell division and when a critical shortness is reached they trigger a DNA damage response that leads to replicative senescence or apoptosis. Telomere shortening is a recognised hallmark of cellular ageing and seems to be also associated with organismal ageing. Telomere length (TL) and the rate of shortening vary across individuals and several studies have found that short telomeres and fast telomere depletion are associated with poor survival and early onset of age related diseases. However, longitudinal studies are needed to better understand the relationship of TL and TL dynamics with longevity measures. Relevant studies on livestock species are largely missing from the literature. In the dairy industry, farmers are forced to cull a considerable percentage of their heifers and cows at a young age due to fertility problems or diseases. As a consequence many replacement heifers have to be reared to maintain a specific herd size. This results in increased costs, consumption of resources, and damage to the environment. Breeding for an improved productive lifespan is difficult because longevity measures are recorded at the end of life and are known to have a low heritability. Therefore, the expected genetic improvement is generally slow, but could be considerably accelerated if an early life heritable biomarker was identified that is predictive of productive lifespan and could be used for animal selection. The question is if TL could be used as such a biomarker. The objectives of this thesis were to 1) develop robust methods to measure average relative leukocyte TL (RLTL) in cattle, 2) examine RLTL dynamics with age at a population as well as at an individual level, 3) estimate genetic parameters and 4) assess the association of RLTL and RLTL dynamics with productive lifespan. A quantitative polymerase chain reaction (qPCR) based assay developed for human studies was adapted to cattle and delivered robust results (repeatability > 80%, coefficient of variation=0.05). Different DNA extraction methods were tested for their effect on RLTL measurements and it was demonstrated that fast silica based DNA extraction methods are suitable for telomere projects which can improve the sample throughput and enable large-scale projects. Subsequently, RLTL in 1328 whole blood samples of 308 Holstein Friesian dairy cows and additionally in 284 whole blood samples of 38 female calves was measured. Repeatability and random regression models were used for the statistical analysis of telomere data. RLTL decreased considerably within the first year of life, but remained relatively stable afterwards at population level. Animals varied significantly in their amount and direction of telomere change. The genetic correlation between consecutive measurements in the same individual weakened with increasing sample interval from r=1 to r=0.69 which indicates that TL in the beginning of life might be under a different genetic control than TL later in life. For the first time in a livestock species we calculated heritability estimates for RLTL which were high (0.32-0.38) and remained constant over life. Long telomeres at birth were not predictive of better productive lifespan. However, animals with long RLTL at the ages of one and five years had a survival advantage. Also, animals that showed less average RLTL attrition over their lives remained in production for longer. TL dynamics differed among individuals and a considerable subset of individuals demonstrated telomere lengthening between consecutive measurements. On average, telomeres tend to shorten early in life and then remain relatively constant. While TL is a heritable trait throughout lifetime, telomere change is not heritable. Short TL at specific ages and telomere attrition over life were associated with poorer productive lifespan.

The primary study aim was to evaluate the latent biological effect of childhood sexual abuse (CSA) on adults by quantifying acquired cytogenetic changes and cortisol levels in identical twins who were discordant (N=22) or concordant (N=2) for a history of CSA. Although the difference scores for cortisol values between discordant identical co-twins were not significantly different from zero, a trend was observed for the twins exposed to intercourse, the most severe form of CSA, to have a blunted cortisol awakening response. Acquired cytogenetic changes were assessed by scoring telomere lengths and somatic cell abnormality frequencies via a cytokinesis-block micronucleus (MN) assay. No significant difference in overall telomere intensity values was observed between co-twins, but chromosome-specific telomere differences were observed in the individuals exposed to intercourse compared to their unabused co-twins ([χ2(45)= 62.88; p= 0.040 and χ2(45)= 73.72; p= 0.004). Specifically, shortened telomeres were observed on the short arms of chromosomes 3, 5, & 6, and long arms of chromosomes 11 & 13. A significant increase in MN frequencies was observed in the abused twins compared to unabused twins (t=2.65; df=16; p=0.009). A significant interaction between micronuclei frequencies and age was also observed, suggesting that the biological effects of stress are cumulative (coefficient [SE] = 0.030 [0.009]; p=0.0006). However, the pattern of chromatin present in MN, which was assessed using spectral karyotyping methodologies, was not limited to the subset of chromosomes with telomeric attrition. In summary, this is the first assessment of acquired chromosomal abnormalities, chromosome-specific telomere lengths and cortisol levels in identical adult twins discordant for exposure to CSA. Given that a portion of biological changes were most pronounced in the intercourse discordant twins, these findings support a possible dose-response relationship with CSA severity. Our data also suggest that the MN assay is a superior tool in assessing the latent effects of stress compared to either cortisol profiling or the measurement of telomere lengths. Collectively, application of the information gained from these studies may allow for novel screening techniques to identify individuals who are most at risk for developing stress-associated disease states.

Transtornos mentais graves são doenças crônicas altamente incapacitantes que geram um alto custo para a sociedade. Indivíduos acometidos por essas doenças apresentam maior morbidade e mortalidade. Dentre elas, a esquizofrenia parece possuir os piores desfechos. Portanto, estudar a esquizofrenia pode trazer contribuições importantes para o entendimento e manejo de transtornos mentais graves como a depressão maior e o transtorno bipolar. Esse trabalho buscou compreender mecanismos fisiopatológicos da esquizofrenia ao longo de quatro artigos que exploram aspectos de funcionamento cognitivo, funcionamento intelectual, de biomarcadores e de estrutura cerebral. O primeiro artigo investigou as alterações de performance de memória em indivíduos com esquizofrenia em estágios iniciais e tardios da doença comparadas ao transtorno bipolar e a sujeitos saudáveis. Os resultados mostraram que indivíduos com esquizofrenia apresentaram precoces prejuízos cognitivos de memória, diferentemente de indivíduos com transtorno bipolar quando comparados a controles. O segundo artigo investigou as influências das performances cognitiva e intelectual em estruturas cerebrais de indivíduos com esquizofrenia comparados a controles saudáveis. Os resultados indicaram que o funcionamento intelectual pré-morbido estava relacionado ao volume de estruturas globais, enquanto o funcionamento cognitivo estava relacionado ao volume e espessura de massa cinzenta cortical, sugerindo influências diferentes e complementares relacionadas a neurodesenvolvimento e neurodegeneração. O terceiro artigo investigou um biomarcador de envelhecimento precoce, um possível mecanismo para a neuroprogressão na esquizofrenia. Os resultados monstraram que indivíduos com esquizofrenia apresentaram encurtamento de telômero quando comparados a controles, mas não houveram diferenças entre o tamanho de telômero de pacientes e seus irmãos não afetados pela doença. Por fim, o quarto artigo buscou investigar a teoria do envelhecimentoa patológico acelerado na esquizofrenia, integrando os achados dos artigos anteriores. Os resultados demonstraram correlações entre comprimento de telômero, níveis de CCL11, performance de memória, volume de massa cinzenta e tempo de doença em indivíduos com esquizofrenia. Esses achados sugerem que a esquizofrenia seria uma doença do neurodesenvolvimento associada a uma carga adicional ao longo do curso da doença que levaria a um envelhecimento patológico precoce. A partir dos achados em esquizofrenia, pode-se ampliar a compreensão de alterações percebidas nas trajetórias de outras psicopatologias. Com adequado entendimento desses mecanismos, será possível o desenvolvimento de novos tratamentos e intervenções mais efetivas e eficazes. / Severe mental disorders are debilitating chronic diseases that have a high cost to society. Individuals affected by these diseases have increased morbidity and mortality. Among them, schizophrenia seems to have the worst outcomes. Therefore, studying schizophrenia may provide important contributions to the understanding and management of severe mental disorders such as major depression and bipolar disorder. The present study aimed to understand the pathophysiological mechanisms of schizophrenia over four articles that explore aspects of cognitive functioning, intellectual functioning, biomarkers and brain structure. The first article investigated changes in memory performance in individuals with schizophrenia in early and late stages of disease compared to bipolar disorder and healthy subjects. The results showed that subjects with schizophrenia had early cognitive deficits of memory, unlike individuals with bipolar disorder compared to controls. The second article investigated influences of cognitive and intellectual performances on brain structures of individuals with schizophrenia compared to healthy controls. The results indicated that premorbid intellectual functioning was related to volume of global structures, while cognitive functioning was related to volume and thickness of cortical gray matter, suggesting different and complementary influences related to neurodevelopment and neurodegeneration. The third article investigated an early aging biomarker, a possible mechanism of neuroprogression in schizophrenia. The results showed that individuals with schizophrenia had shortened telomeres when compared to controls, but there were no differences between the telometer length of patients and their siblings not affected by the disease. Finally, the fourth article sought to investigate the theory of pathological accelerated aging in schizophrenia, integrating the findings of the previous articles. The results demonstrated correlations between telometer length, CCL11 levels, memory performance, gray matter volume and illness duration in individuals with schizophrenia. These findings suggest that schizophrenia is a neurodevelopmental disorder associated with an additional burden over the course of the disease that leads to a pathological accelerated agig.

Telomeres are protective structures at the end of our chromosomes, composed of multiple repeats of the DNA sequence TTAGGG. They are essential for maintaining chromosomal stability by preventing damage and degradation of the chromosome ends. Telomeres are normally shortened with each cell division until a critical length is reached, at which stage cell cycle arrest is induced. Telomere shortening can be prevented in the presence of the telomere-­‐elongating enzyme telomerase. Telomerase is expressed during embryogenesis and in certain normal cell types, but most somatic cells exhibit undetectable levels of telomerase activity. In contrast, most cancer cells express telomerase enabling them to proliferate indefinitely. There is a search for reliable molecular markers that can be used to help predict cancer risk and outcome. The interest of investigating telomere length as a potential biomarker in malignancy has grown rapidly, and both tumors and normal tissues have been in focus for telomere length measurements. In this thesis, telomere length was investigated in breast cancer patients and in patients with renal cell carcinoma (RCC). The breast cancer patients were found to have significantly longer mean telomere length in peripheral blood cells (i.e. immune cells) compared to a tumor-­‐free control group. Moreover, patients with the longest blood telomere length had a significantly worse outcome compared to patients with shorter blood telomeres. In a patient group with clear cell RCC, telomere length was investigated in peripheral blood cells, in tumors and in corresponding kidney cortex. Again, patients with the longest blood telomere length had a significantly worse prognosis compared to those with shorter blood telomeres. In contrast, telomere length in tumor and kidney cortex tissues did not predict outcome per se. Immunological components may play a role in telomere length dynamics as well as in cancer development. We aimed to investigate a possible association between telomere length and certain immunological parameters, including various cytokines and peripheral levels of a blood cell type with suppressor function [regulatory T cells (Tregs)]. In our patients with clear cell RCC, three cytokines correlated significantly with tumor telomere length, but not with telomere length in peripheral blood cells. In a separate patient group with various RCC tumors, blood telomere length correlated positively with the amount of Tregs. It might be speculated that a subset of patients with long blood telomeres has a less efficient immune response due to high Treg levels, contributing to a worse prognosis. Another aim of this thesis was to explore telomere length changes over time. Evaluation of blood samples collected at a 6-­‐month interval from 50 individuals, showed that half of the participants experienced a decline in mean telomere length during the time period. This group had longer telomere length at baseline compared to those who demonstrated increased/stable telomere length. In a separate group of five blood donors, a remarkable drop in telomere length was detected in one donor over a 6-­‐month period, whereas the other donors exhibited only small fluctuations in telomere length. In conclusion, the results of this thesis indicate that blood telomere length has potential to act as an independent prognostic marker in malignancy. Adding to the complexity is the fact that changes in blood telomere length might occur within relatively short time spans, indicating that telomere length is a dynamic character.

Telomeres are repetitive TTAGGG structures ending each chromosome and thereby protecting its integrity. Due to the end-replication problem, telomeres shorten with each cell division. When reaching a critical telomere length (TL), the cells stop dividing and enter replicative senescence. It has been speculated that telomeres might regulate lifespan at the organism level but this hypothesis is controversial. However, telomeres in human blood cells do shorten with increasing age. Telomerase is an enzyme capable of lengthen telomeres. It consists of a catalytic subunit, hTERT, and a RNA template, hTR. Telomerase is active in germ cells, stem cells, activated lymphocytes and highly proliferating epithelial cells while no activity is found in other somatic cells. One step in order to produce a tumour mass is that cancer cells need to have a limitless replicative potential and this can be achieved by activating telomerase. Most tumour cells express telomerase activity and hence, the enzyme is an interesting target for cancer therapy. Telomere length is in part inherited. Two separate family cohorts were investigated to elucidate the inheritance pattern and a strong paternal inheritance was observed. In the larger, multifamily cohort spanning up to four generations, a weak correlation between the TL of the mother and the child was also found, as well as a significant correlation between grandparent-grandchild pairs. Interestingly, the heritable impact diminished with increasing age, indicating than non-heritable factors might influence TL during life. A functional T to C transition polymorphism in the hTERT promoter was previously reported, showing that the -1327C/C genotype was correlated with shorter TL compared to the alternative genotypes in healthy individuals and in coronary artery disease patients. When investigating 226 myocardial infarction patients and 444 controls separately, no differences were observed regarding mean TL or increased attrition rate between the different genotypes. TL in blood cells is shown to be altered in patients with certain types of solid tumours. In our breast cancer cohort, TL was a strong prognostic marker. Short telomeres were associated with increased survival, especially in young patients and in those with advanced tumours. It has been speculated that cancer patients might have a faster telomere attrition rate than controls but this has not been experimentally proven. Two blood samples from the same individual taken with 9-11 years interval was investigated. Some were diagnosed with a malignancy after the second blood draw. When comparing patients with controls, telomere attrition rate was not correlated to future tumour development. About one third of the individuals elongated their telomeres over a decade and the individual telomere attrition rate was telomere length dependent, showing an inverse correlation to TL at a highly significant level. This strongly suggests that the TL maintenance mechanism shown to provide protection for short telomeres in vitro is important also in human cells in vivo.