Life Sciences Commons^™

Multi-Omic And Single-Cell Characterization Of A 3d Skin-Like Tissue Model Of Systemic Sclerosis With A Focus On Epigenetics, Tamar R. Abel

Dartmouth College Ph.D Dissertations

Systemic sclerosis (SSc) is a rare fibrotic autoimmune disease with high mortality and limited FDA approved therapies. Clinical concordance among twins is low; however, a modest familial heritability implicates complex interactions between polygenic risk alleles and the environment as causal – interactions that are mediated by epigenetics. Although mechanisms of tissue fibrosis have been successfully identified and targeted in established in vitro models for SSc, these molecules have ultimately failed in clinical trials. This likely reflects the lack of a reliable in vitro model that faithfully recapitulates the disease.

I utilized a 3D skin-like tissue model to study SSc skin …

Novel Mechanistic Insight Into Ciliary Regulation: Old Pathways Yield New Mechanisms, Larissa L. Dougherty

Dartmouth College Ph.D Dissertations

Cilia are structures present on most eukaryotic cells which provide important signaling and motile components to cells from early development to fully differentiated and matured cells. Regulation of these structures is critical to proper functioning of the cell and is known to be tied to the cell cycle. Preparation for ciliary assembly following cell cycle exit and ciliary disassembly following cell cycle reentry requires components throughout the cell body and within the cilium to facilitate this process. Here I identify how the cell adapts to ensure modifications to cilia occur for assembly or disassembly using the model organism Chlamydomonas reinhardtii. …

Mitochondrial Division: Synergizing In Mitochondrial Divisome, Ao Liu

Dartmouth College Ph.D Dissertations

Mitochondria are the energy factories of the cell. The dynamic nature of cells demands routine changes in mitochondrial morphology by fusion and division. The dynamin GTPase Drp1 is a central mitochondrial division protein, driving constriction of the outer mitochondrial membrane via oligomerization. At least four regulatory factors control Drp1 activity on the outer mitochondrial membrane (OMM): 1) receptor proteins (Mff, MiD49, MiD51, and Fis1); 2) actin filaments; 3) the mitochondrial phospholipid cardiolipin (CL); and 4) Drp1 post-translational modifications, of which two phosphorylation sites (S579 and S600) are the most well studied. However, the molecular mechanism of how these factors work …

Visual Ecology: Sensing The Social World, Elliott P. Steele

Dartmouth College Ph.D Dissertations

Social animals, and even solitary animals, must at different points in their life actively search for and locate conspecifics within their environment, using one or multiple senses. One sense in particular, vision, can provide some of the most spatially detailed and temporally rapid information about the environment. Yet, the surrounding environment can also severely challenge animals’ abilities to find fellow conspecifics. Furthermore, once an animal successfully locates conspecifics, it must then make key decisions about when and where to enter group formations, decisions which may strongly impact their access to socially-acquired resources. Even after an animal has positioned itself within …

Ecology And Evolution Of Social Information Use, Clare T. M. Doherty

Dartmouth College Ph.D Dissertations

Sociality is a strategy many animals employ to cope with their environments, enabling them to survive and reproduce more successfully than would otherwise be possible. When navigating their environments and making decisions, social individuals often use information provided by conspecifics (in the form of social cues and signals), thereby increasing the scope and reliability of the information they can gather. However, social information use may be influenced by many factors, including key differences in context across the physical and social environment. My thesis asks and answers a series of questions regarding the trade-offs in social information use across different contexts, …

Regulation Of Tissue Mechanics And Adherens Junctions By Small Gtpase Rhoa During Drosophila Embryogenesis, Hanqing Guo

Dartmouth College Ph.D Dissertations

Actomyosin contractility plays an important role at both the cell and tissue level during developments. In this study, we developed an optogenetic tool that can acutely inhibit actomyosin contractility by targeting its main activator Rho1. This optogenetic tool can achieve myosin inhibition within one minute and thus enable further dissection of actomyosin function in development. In my first two projects, I used Drosophila mesoderm invagination (also known as ventral furrow formation) as a model to study epithelial folding, a fundamental mechanism for constructing complex 3D tissues. Apical constriction mediated by actomyosin contractility is a common mechanism for epithelial folding. However, …

Mechanisms And Roles Of Dynamic Actin Assembly Around Dysfunctional Mitochondria, Tak Shun Fung

Dartmouth College Ph.D Dissertations

Possessing the ability to efficiently generate ATP required to sustain cellular functions, mitochondria are often considered the ‘powerhouses of the cell’. However, our understanding of mitochondria in cell biology was further expanded when we recognized that communication between this unique organelle and the rest of the cell regulates cellular bioenergetics, metabolism and signaling processes such as mitophagy and apoptosis. Here, I investigate signaling between mitochondria and the actin cytoskeleton, and how this signaling regulates mitochondrial dynamics and cellular function. Specifically, I find that, upon mitochondrial dysfunction, actin polymerizes rapidly around the dysfunctional organelle, which we term ‘acute damage-induced actin’ (ADA). …

Consequences Of Biofilm Architecture On Vibrio Cholerae Ecology And Life History, Benjamin Ray Wucher

Dartmouth College Ph.D Dissertations

The diversity of microbes and the environments they inhabit are staggering. In many of these environments, bacteria have evolved to form sessile surface attached communities called biofilms. These biofilms have wide reaching impacts from importance in global carbon cycling, to persistent catheter infections, to biofouling and wastewater treatment. While many species of microbes form biofilms to survive in their environment, the architectures of these structures vary widely between organisms. Even though a great deal of work has been done to understand bacterial communities and their functions, little work has examined how the spatial aspects of biofilm architecture can affect the …

The Roles Of Epithelial–Mesenchymal Plasticity In Tumor Heterogeneity, Metastasis, And Patient Survival In Breast Cancer, Meredith Septer Brown

Dartmouth College Ph.D Dissertations

The Epithelial-to-Mesenchymal transition, a critical cellular process in development, is frequently co-opted by solid tumors to promote invasion and metastasis. In particular, the hybrid or intermediate EMT state, possessing both epithelial and mesenchymal characteristics, is associated with increased cancer stemness and plasticity. Similarly, intra-tumoral heterogeneity in solid tumors, in particular breast cancer, is associated with poor prognosis, tumor growth, proliferation, drug resistance, and metastasis. We sought to understand the link between the generation of intra-tumoral heterogeneity and the intermediate EMT state and their impact on tumor progression and patient prognosis. As part of my thesis work, I developed a model …

Novel Functions For Arp2/3 Complex-Mediated Actin Networks Discovered In Chlamydomonas Reinhardtii, Brae M. Bigge

Dartmouth College Ph.D Dissertations

Chlamydomonas reinhardtii, a unicellular member of the Chlorophyta or Green Algae phylum, has been used for decades as a model for ciliary studies. Using this tool, previous work from our lab found a role for actin in ciliary assembly and maintenance. However, while the microtubule-based, membrane-ensheathed cilia of Chlamydomonas are highly conserved in relation to mammalian cells, the actin cytoskeleton is not as simple. Chlamydomonas contains two actin genes: IDA5, a conventional actin, and NAP1, a divergent actin. Here, we find that despite the divergence of NAP1, it is still able to interact with the actin nucleator, the Arp2/3 complex. …

Spatial Representation In Postrhinal Cortex, Patrick Lachance

Dartmouth College Ph.D Dissertations

Animals rely on a variety of internal and external cues to orient themselves when navigating their environments and determining their current spatial context. Information regarding these cues enters the brain from the navigator’s first-person perspective. Information of this type is considered to be egocentric, or self-centered. However, decades of behavioral, electrophysiological, and imaging research suggest that the brain contains a rich collection of spatial representations that are unrestricted by the animal’s first-person perspective, and instead are defined relative to the surrounding environment. These representations are considered allocentric, or world-centered. Despite an abundance of promising modeling work, the specific mechanisms by …

The Role Of L-Alanine Signaling In Aspergillus Fumigatus Biofilm Adherence, Carbon Catabolism, And Echinocandin Susceptibility., Joshua D. Kerkaert

Dartmouth College Ph.D Dissertations

Aspergillus fumigatus is a saprophytic filamentous fungus that participates in environmental carbon and nitrogen cycles through the degradation of complex organic substrates. In addition to its ecological role, A. fumigatus is the primary causative agent of a spectrum of diseases depending on the immune status of the individual, the most lethal of which is invasive aspergillosis (IA). Treatment strategies for IA are limited and far too frequently fail. Despite the high rates of treatment failure, antifungal resistance remains relatively low, albeit rising at a concerning rate. Insights into this discrepancy between the rate of treatment failure and the rate of …

Deciphering Phosphoprotein Phosphatase Signaling Networks Using Proteomics Approaches, Brooke Brauer

Dartmouth College Ph.D Dissertations

Protein phosphorylation is a highly regulated mechanism of cell signaling control and its deregulation is implicated in disease. The kinases that catalyze the addition of phosphate groups onto their substrate proteins have been well studied, their signaling pathways mapped, and their effects on cell and organismal health observed. Knowledge of the phosphatases that reverse the reaction only recently began to come into focus. Phosphoprotein phosphatases (PPPs), long thought to be housekeeping enzymes, are now known to be exquisitely specific towards their substrates, but the exact nature of phosphatase regulation—both upstream and downstream of the phosphatase—is unclear.

PPPs recognize substrates through …

Genetic Heterogeneity In Schizophrenia And Contribution Of Context To Vowel Recognition, Eva Childers

Dartmouth College Ph.D Dissertations

This dissertation is composed of three chapters that address two distinct topics. Chapter 2 addresses the use of consonantal context in vowel perception. Previous studies have demonstrated that context is important for vowel identification, however, this effect may be an artifact of stimuli production. To address this potential confound, we used stimuli extracted from an audiobook and asked subjects to identify vowels encased in consonants and isolated by themselves. We show that subjects had improved vowel identification when the vowel is presented with a consonantal framing, suggesting there is information contained in the surrounding context that is important for phoneme …

Deciphering Taxa-Function Relationships In Population-Level Studies Of Human Gut Microbiomes, Quang P. Nguyen

Dartmouth College Ph.D Dissertations

The human gut microbiome is a complex and dynamic ecosystem, featuring a multitude of microbes all interacting with their hosts in an elaborate manner. Even though this exchange is often mediated through microbial metabolic and functional outputs, such as the production of certain metabolites, environmental exposures, and host lifestyle are highly influential in shaping the presence of microbial species irrespective of their individual roles. As such, a comprehensive understanding of the microbiome requires researchers to examine the relationship between taxonomic abundance and function simultaneously. Assessing microbial contributions to important ecosystem services can enable identification of robust functions supported by a …

Microbial Sociality In Biofilms, Swetha Kasetty

Dartmouth College Ph.D Dissertations

Biofilms are communities of microorganisms attached to surfaces through various self-secreted matrix materials. Biofilms are dynamic communities with extensive interactions between their residents. Microorganisms compete, cooperate, and communicate with each other in biofilms. These ecological interactions determine the emergence or loss of strains/species and are critical in the formation and proliferation of biofilms. Furthermore, most natural biofilms are formed by multiple microbial species and strains and these interactions within the biofilm dramatically influence community composition and structure over time. Microbial interactions also influence clinically relevant outcomes such as antibiotic resistance and host virulence. In this thesis we explore the potential …

Cysteine Metallochemistry And Metal Binding: Quantification Of The Thermodynamic Foundations Of Cellular Homeostasis, Matthew R. Mehlenbacher

Dartmouth College Ph.D Dissertations

Metals are required for life. Many metalloproteins contain cysteine in their metal-binding site (MBS) and cysteines are unique in that they are reactive, and strongly bind certain metals, which aid in metal selectivity and specificity. Using isothermal titration calorimetry (ITC), the thermodynamic foundation for metal binding, cellular protection, and transcriptional regulation, which all utilize cysteines in their MBS, are quantified.

In bacteria there are metalloprotein pathways that actively uptake mercury, which are regulated by the metalloregulatory protein MerR. MerR de-represses the transcription of these mer proteins in a metal-dependent manner. Using ITC, the thermodynamic foundation of the negative allosteric coupling …

Links Between Electrophilic Stress And Antifungal Resistance In Pathogenic Candida Species, Amy R. Biermann

Dartmouth College Ph.D Dissertations

Collectively, Candida species are the most prevalent cause of both superficial and invasive fungal infections worldwide. Invasive Candida infections have a high mortality rate and predominantly affect individuals with underlying diseases, such as diabetes, HIV, or cancer. Unfortunately, many invasive Candida infections are recalcitrant to antifungal treatment, while intrinsically multidrug-resistant pathogens, like Candida auris, are increasing in prevalence. Although the canonical mechanisms of antifungal resistance in Candida species are well established, i.e., overexpression of efflux pumps and overexpression of or mutations in genes encoding drug targets, factors affecting the natural evolution and regulation of resistance mechanisms remain poorly understood. …

Beyond Homogenous Decision-Making Models: Role Of Brain Areas Interaction And Heterogeneity, Mohsen Rakhshan

Dartmouth College Ph.D Dissertations

Most existing mechanistic models of decision making consider microcircuits within a homogenous brain area and fail to consider the distributed nature of this cognitive process. More specifically, these models do not take into account: a) how different types of information might influence choice in different stages of the process; and b) how interactions and the heterogeneity between circuits across different brain areas might be essential for the process. In this dissertation, I present a combination of experimental and modeling approaches to study distributed nature of the decision-making by examining both the distributed processing of information as well as the interaction …

In Situ Vaccination Using Plant Viral Particles As A Novel Approach For Cancer Immunotherapy, Chenkai Mao

Dartmouth College Ph.D Dissertations

Non-infectious to mammals, genetically, and physically stable plant virus-based nanoparticles have been increasingly studied as a novel platform for cancer vaccine and adjuvants due to their immunogenic properties. Our prior work showed that, empty cowpea mosaic virus (eCPMV), a plant virus-like particle (VLP) that is composed of a separated capsid, is an excellent in situ vaccine (ISV) for cancer immunotherapy; however, little is known about the mechanisms linked to their activities regarding recognition and antigen presentation.

In the present studies, we report cowpea mosaic virus (CPMV) and RNA-lacking CPMV (empty, eCPMV) are both toll like receptor (TLR) agonists for TLRs …

Profiling Durable Anti-Tumor Memory T Cell Responses In Long-Term Melanoma Survivors, Jichang Han

Dartmouth College Ph.D Dissertations

While T-cell responses to cancer immunotherapy have been avidly studied,

long-lived memory has been poorly characterized. Of melanoma patients who

receive immunotherapy, long-term survivors are frequently found to develop

melanoma-associated vitiligo. Our prior work showed in a preclinical model that

vitiligo skin sustained a long-lived CD8+ resident memory T cell (TRM) population,

playing key roles in perpetuating anti-tumor immunity. However, the characteristics

and longevity of these memory T cells in melanoma survivors have not been defined.

In the present studies, we probed both the CD8+ and CD4+ T cell responses,

focusing on memory T cell responses in a cohort of …

Response And Molecular Control Of Cd8 T Cells During Infection And Cancer, Nicholas K. Preiss Dr.

Dartmouth College Ph.D Dissertations

CD8 T cells are potent immune effector cells capable of vast clonal expansion and clearance of infected or cancerous cells. After control of the pathogenic insult, CD8 T cells develop into quiescent, long-lived memory populations that are poised to mediate rapid protection upon reencounter with cognate antigen. These properties make control of CD8 T cell responses a highly desirable outcome of vaccine strategies and immunotherapy. Therefore, understanding how the effector function and memory differentiation of CD8 T cells are controlled at a molecular level is of great importance. In the context of infection with gammaherpesviruses (γHV), which form a latent …

Discretized Geometric Approaches To The Analysis Of Protein Structures, John Holland

Dartmouth College Ph.D Dissertations

Proteins play crucial roles in a variety of biological processes. While we know that their amino acid sequence determines their structure, which in turn determines their function, we do not know why particular sequences fold into particular structures. My work focuses on discretized geometric descriptions of protein structure—conceptualizing native structure space as composed of mostly discrete, geometrically defined fragments—to better understand the patterns underlying why particular sequence elements correspond to particular structure elements. This discretized geometric approach is applied to multiple levels of protein structure, from conceptualizing contacts between residues as interactions between discrete structural elements to treating protein structures …

Modeling The Effect Of Igg Subclasses And Specificity In The Translocation Of Monoclonal Antibodies Across The Placental Barrier, Sayuri Tais Miyamoto Magnabosco

ENGS 88 Honors Thesis (AB Students)

Infections are responsible for over half a million neonatal deaths every year (Lawn et al., 2014). Thus, there is huge interest in leveraging maternal immunization against infectious diseases to grant fetal protection during its development through the vertical transferring of IgG antibodies, the only Ig subclass that can significantly cross the placental barrier. Studies about vertical immunization rely on in-vitro models to extrapolate physiological conditions of the human placenta. The BeWo Transwell model (Bode et al., 2006) presents itself as a reliable model to mimic the transplacental transport mechanism of antibodies (Ellinger et al., 1999; Poulsen et al., 2009) …

Flash Radiotherapy: Skin Pigmentation As A Non-Invasive Indicator For Radiation-Induced Damage, Brady Mccallister

ENGS 88 Honors Thesis (AB Students)

FLASH ultra-high dose rate radiotherapy (RT) is one of the most rapidly growing subfields of radiation oncology today due to its potential to increase the limits of the therapeutic ratio. The FLASH effect, which includes heightened normal tissue sparing paired with iso-effective tumor cell killing, has been literature documented, in a limited manner, in rodent models, a few large animals, and one clinical patients.

A porcine-based experiment was conducted to test the effects of FLASH RT on normal tissue compared to conventional (CONV) RT. A clinical linear accelerator (LINAC) was reversibly converted to be capable of FLASH RT. A female …

Application Of Cycle-By-Cycle Analysis To Eeg Data From Individuals With Phelan-Mcdermid Syndrome, Naomi Miller

ENGS 88 Honors Thesis (AB Students)

This study aimed to analyze a novel method of processing data from electroencephalography (EEG) recordings, which implements time-domain cycle-by-cycle analysis. This "bycycle" method, developed by the Cole & Voytek laboratory, was implemented on a EEG dataset of children with and without Phelan-McDermid Syndrome in the hopes of uncovering network-level explanations for the genetic disorder. A supplemental Python pipeline was developed to organize and visualize the data. This led to the discovery of group-level differences in measures of cycle symmetry in alpha band waves over the sensorimotor electrodes. Through the same pipeline, the bycycle tool was validated as a sound EEG …

Nmda Receptors Enhance The Fidelity Of Synaptic Integration, Chenguang Li, Allan Gulledge

Dartmouth Scholarship

Excitatory synaptic transmission in many neurons is mediated by two coexpressed ionotropic glutamate receptor subtypes, AMPA and NMDA receptors, that differ in kinetics, ion selectivity, and voltage-sensitivity. AMPA receptors have fast kinetics and are voltage-insensitive, while NMDA receptors have slower kinetics and increased conductance at depolarized membrane potentials. Here, we report that the voltage dependency and kinetics of NMDA receptors act synergistically to stabilize synaptic integration of EPSPs across spatial and volt- age domains. Simulations of synaptic integration in simplified and morphologically realistic dendritic trees re- vealed that the combined presence of AMPA and NMDA conductances reduce the variability of …

A Multi-Resolution Graph Convolution Network For Contiguous Epitope Prediction, Lisa Oh

Dartmouth College Master’s Theses

Computational methods for predicting binding interfaces between antigens and antibodies (epitopes and paratopes) are faster and cheaper than traditional experimental structure determination methods. A sufficiently reliable computational predictor that could scale to large sets of available antibody sequence data could thus inform and expedite many biomedical pursuits, such as better understanding immune responses to vaccination and natural infection and developing better drugs and vaccines. However, current state-of-the-art predictors produce discontiguous predictions, e.g., predicting the epitope in many different spots on an antigen, even though in reality they typically comprise a single localized region. We seek to produce contiguous predicted epitopes, …

Towards Sustainable Aquafeeds: Evaluating Substitution Of Fishmeal With Lipid-Extracted Microalgal Co-Product (Nannochloropsis Oculata) In Diets Of Juvenile Nile Tilapia (Oreochromis Niloticus), Pallab K. Sarker, Anne R. Kapuscinski, Ashley Y. Bae, Emily Donaldson, Devin S. Fitzgerald, Oliver F. Edelson

Dartmouth Scholarship

Microalgae companies increasingly seek markets for defatted biomass that is left over after extracting omega-3 rich oil for human nutraceuticals and crude oil for fuels. Such a protein-rich co-product is a promising alternative to unsustainably sourced fishmeal in aquaculture diets. We report the first evaluation of co-product of the marine microalga Nannochloropsis oculata(N. oculata co-product) for replacing fishmeal in diets of Nile tilapia, a globally important aquaculture species. We conducted a nutrient digestibility experiment with N. oculata dried whole cells and N. oculata co-product, followed by an 84-day nutritional feeding experiment with N. oculata co-product. N. oculata co-product, more nutrient-dense …

Idiosyncratic, Retinotopic Bias In Face Identification Modulated By Familiarity, Matteo Visconti Di Oleggio Castello, Morgan Taylor, Patrick Cavanagh, Maria Ida Gobbini

Dartmouth Scholarship

The perception of gender and age of unfamiliar faces is reported to vary idiosyncratically across retinal locations such that, for example, the same androgynous face may appear to be male at one location but female at another. Here we test spatial heterogeneity for the recognition of the identity of personally familiar faces in human participants. We found idiosyncratic biases that were stable within participants and that varied more across locations for low as compared to high familiar faces. These data suggest that like face gender and age, face identity is processed, in part, by independent populations of neurons monitoring restricted …