Kloth, Alexander

• Postdoc., Neuroscience, University of North Carolina, Chapel Hill, NC 

• Ph.D., Molecular Biology and Neuroscience, Princeton University, Princeton, NJ 

• M.A., Molecular Biology, Princeton University, Princeton, NJ 

• Postbacc., Mental Health, National Institutes of Health, Bethesda, MD 

• B.S.E., Biomedical Engineering, Duke University, Durham, NC

Autism spectrum disorder (ASD) is a neurodevelopmental disorder marked by socio-communicative deficits, repetitive behaviors, systemic interests, and other neurological and cognitive symptoms that have a wide range of severity. It is estimated that ~2.3% of children born today will receive an ASD diagnosis, with males estimated to be about three times as likely to receive an ASD diagnosis. Because of the high prevalence of ASD, decades of preclinical research have investigated the disorder's complex etiology and its manifestation in the brain, with the goal of uncovering effective therapeutic strategies. Despite these efforts, no treatments addressing ASD's core features have been successfully translated to the clinic.

A long-term goal of our laboratory is to determine whether treatments with neurotrophic actions-including environmental enrichment and pharmacological agents-might one day lead to successful therapies for ASD. One possible strategy that has recently attracted our attention involves erythropoietin (EPO) and its derivatives. EPO is an endogenous cytokine that has potent, long-lasting effects on cell survival, neuroplasticity, and neurogenesis, making it a candidate for treating a host of neuropsychiatric disorders, including ASD. However, EPO's potential has been limited by it severe hematological side effects. To overcome this hurdle, researchers have engineered non-hematopoietic derivatives that appear to have similar actions as EPO and appear to ameliorate disease features in model mice. For instance, we have recently shown that carbamoylated EPO (CEPO), which rescues anxiety and depression-related behavior in the BALB/c mouse model, also restores social approach behavior in these mice. However, the degree to which CEPO rescues social and other ASD-related behaviors, the mechanisms underlying these potential behavioral changes, and the applicability of these findings to other ASD models all remain unclear.

The goal of the proposed project is to more broadly assess the effects of CEPO on ASD-related behaviors and examine the underlying neurobiological correlates of these effects. To do so, we will first employ a rigorously characterized idiopathic ASD mouse model, the BTBR T+Itpr3tfIJ (BTBR) mouse (Aims 1 & 2). BTBR mice show a broad suite of ASD-related behavior deficits and demonstrated decreased neurotrophic factor levels, diminished adult neurogenesis, aberrant neuronal morphology, and abnormal glial activity in the hippocampus, which are all potentially corrected by CEPO. We hypothesize the CEPO administration will rescue ASD-related social behaviors n BTBR mouse in part by correcting these cellular and molecular defects. We will also examine the necessity of the endogenous erythropoietin receptor (EpoR), on which CEPO is thought to act, for ASD-related behaviors (Aim 3). We hypothesize that a conditional EpoR knockout will recapitulate the effects corrected in BTBR mice. To test these hypotheses, we will complete the following aims:

Aim 1. Determine the degree to which CEPO administration rescues ASD-related behaviors in BTBR mice. BTBR mice show a range of social deficits, deficient ultrasonic vocalizations in social situations, and other nonsocial ASD-relevant phenotypes. We will determine whether  CEPO administration corrects the traditional suite of ASD-related behaviors in BTBR mice, including social approach, exploration in the open field, motor function on the rotarod, and grooming and burrowing behavior. In a further experiment, we will examine whether these corrections extend to social interactions and vocalizations in a more naturalistic setting by using artificial intelligence methods for tracking animal behavior.

Aim 2. Examine neurobiological correlates of CEPO affects in the hippocampus in BTBR mice. Behavioral deficits in BTBR  mice have been associated with molecular, cellular, and neural circuits effects, including aberrant circuit activation, cell morphology, and glial activation, reduced neurotrophic signaling, disrupted neurogenesis, and disrupted neurotransmission. Focusing on the hippocampus-a common region of interest in BTBR mouse model-we will use a combination of immunohistochemistry, histology, and qRT-PCR to determine what effects CEPO has on activation and cellular makeup of the hippocampus, signaling mechanisms, and the survival of newborn neurons, and how these effects correlate to the findings in Aim 1.

Aim 3. Investigate the necessity of hippocampal EpoR expression for ASD-related phenotypes. CEPO is thought to act by binding to the endogenous EpoR receptor, which is highly expressed in the hippocampus. We will test whether a CaMKII-conditional knockout of EpoR shows similar behavioral  deficits and patterns of neuronal activation and neurotrophic signaling as those rescued by CEPO in BTBR mice.

This project will provide undergraduate students to learn key skills in behavior, histology, immunohistochemistry, and molecular biology - all bedrock techniques in neuroscience - while exposing them to artificial intelligence, microscopy, and animal modeling techniques common in 21st century neuroscience.