About

Robert Spitale is the Founding Associate Dean of Research and a Professor at the School of Pharmacy & Pharmaceutical Sciences at the University of California, Irvine. He earned his Ph.D. in Chemistry from the University of Rochester in 2009. His research focuses on understanding how RNA molecules orchestrate gene expression pathways and contribute to disease pathologies. His laboratory aims to develop biochemical technologies to study RNA inside living cells, addressing important questions about RNA structure and function on a systems level. Spitale's work also emphasizes inspiring learning and educating students, postdoctoral fellows, and clinician-scientists to become leaders in academic and industrial pharmacology, as well as translational sciences. Prior to his current position, he was a Postdoctoral Fellow at Stanford University from 2010 to 2014, working in the laboratory of Howard Y. Chang.

Research topics

• Cell biology
• Biology
• Computational biology
• Biochemistry
• Genetics
• Neuroscience
• Nanotechnology
• Medicine
• Pathology
• Immunology
• Cancer research
• Chemistry

Selected publications

• Fluid and Neuroimaging Biomarkers in Microgliopathy Colony‐Stimulating Factor‐1 Receptor‐Related Disorders

  Annals of Clinical and Translational Neurology · 2026-01-12 · 1 citations

  articleOpen access

  OBJECTIVE: This study aims to identify both fluid and neuroimaging biomarkers for CSF1R-RD that can inform the optimal timing of treatment administration to maximize therapeutic benefit, while also providing sensitive quantitative measurements to monitor disease progression. METHODS: Our study compared neuroimaging and fluid (plasma and cerebrospinal fluid (CSF)) biomarkers across three distinct populations: asymptomatic CSF1R pathogenic variant carriers (N = 14), symptomatic CSF1R pathogenic variant carriers (N = 17), and healthy controls (N = 30). We evaluated biomarker correlations with both an established (Montreal Cognitive Assessment (MoCA)) and a novel (CSF1R Clinical Severity Score (CCSS)) clinical diagnostic scale to investigate potential clinical utility. Additionally, we tested the relationship between select biomarkers and cortical thickness using 3D T1-weighted MPRAGE scans, providing a highly valuable physiological component to our analyses. RESULTS: Our results demonstrate that while plasma glial fibrillary acidic protein (GFAP) displays a high sensitivity for distinguishing early-stage CSF1R-RD patients from healthy controls, plasma neurofilament light chain (NfL) is more effective for tracking disease progression following the onset of symptoms. INTERPRETATION: Overall, our study provides evidence for plasma NfL and GFAP as valuable biomarkers of earliest symptom onset and disease progression for CSF1R-RD.

  PublisherOA PDFDOI

• The Need for Staging System in Colony Stimulating Factor 1 Receptor‐Related Disorder

  Movement Disorders · 2026-02-27

  article

  Data sharing not applicable to this article as no datasets were generated or analysed during the current study. Data S1. Supporting Information. Video S1. The video shows a 38-year-old woman carrying a CSF1R c.1897G>A (p.Glu633Lys) variant. She is in a very early stage of the disease, presenting with mild but noticeable lower-limb ataxia and pyramidal signs that do not significantly interfere with daily functioning. These findings correspond to a motor score of 2. The patient reports depressive and anxiety symptoms; however, at this stage, a causal relationship with CSF1R-RD cannot be clearly established. These symptoms are of moderate severity and do not impair her activities of daily living. Accordingly, she receives 2 points on the neurocognitive scale. Overall, the patient scores 2 on both the motor and cognitive scales, corresponding to stage 2 (early symptomatic) CSF1R-RD. Video S2. The patient is a 44-year-old man carrying the CSF1R c.2375C>A (p.Ala792Asp) variant, with an age of onset of 35 years. He underwent HSCT at age 37 years-old at stage 2 (Neurocognitive – 1, Motor – 2). He presents with pronounced motor symptoms requiring bilateral assistance for ambulation, corresponding to Stage 3 on the motor scale. He also exhibits mood disturbances and subjective cognitive impairment, with a MoCA score of 26/30, corresponding to stage 2 on the neurocognitive scale. The overall disease stage is therefore Stage 3, consistent with mild CSF1R-RD. Video S3. The video shows a 25-year-old female with symptom onset at age 23-years -old. carrying the CSF1R c.670 T>A (p.Cys224Ser) variant. She presents with evident motor symptoms and gait impairment requiring the use of a walker. Symptoms are more pronounced on the right side and include marked spasticity, dystonic posturing of the right limbs, and an alien limb phenomenon, along with clear pyramidal signs such as ancle clonus and brisk deep tendon reflexes. Accordingly, she would be classified as Stage 3 on the motor scale. Despite these motor impairments, the patient remains professionally active, and her MoCA score is 27/30, corresponding to stage 2 in the neurocognitive domain. The overall disease stage is therefore Stage 3, consistent with mild CSF1R-RD. Video S4. The video presents a 45-year-old man diagnosed with CSF1R-RD caused by the c.2442 + 1G>A variant in the CSF1R gene. The patient retains relatively good gross motor function and is able to walk independently; however, he has difficulties with fine motor movements, placing him at Stage 3 on the motor axis. In contrast, significant cognitive impairment necessitates substantial assistance from family members. Although he can perform simple tasks, he requires supervision and encouragement, corresponding to stage 4 on the neurocognitive axis. Overall, his condition is classified as stage 4 (moderate) CSF1R-RD. Video S5. The video shows a 29-year-old female carrying a CSF1R c.2442 + 2 T>C variant, with disease onset at 26 years of age. The patient exhibits significant motor symptoms yet is able to ambulate with assistance. Her overall functional status is largely determined by neurocognitive impairment. She is unable to care for herself, requires total assistance in activities of daily living. She is completely mute. Her MoCA score is 0. The video demonstrates that she can follow only simple commands; for example, she does not understand the command to walk backward. Her score in the neurocognitive domain is 5, contributing to an overall stage of 5, consistent with severe CSF1R-RD. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

  PublisherDOI

• The path to clinical application of human microglia transplantation for the treatment of CSF1R-related disorder

  Expert Review of Neurotherapeutics · 2026-05-12

  article

  INTRODUCTION: gene, leading to dysfunctional microglia. The disease presents with diverse, progressive neurological symptoms, including motor and cognitive decline, and has a poor prognosis. AREA COVERED: This review critically examines the emerging therapeutic landscape for CSF1R-RD, with a primary focus on the disease-modifying potential of Hematopoietic Stem Cell Transplantation (HSCT). It synthesizes data from cohort studies and case reports, analyzing outcomes, neuroradiological changes, and the safety profile of HSCT. Furthermore, the review discusses the utility of advanced neuroimaging and fluid biomarkers, such as Neurofilament light chain (NfL) and Glial fibrillary acidic protein (GFAP), which are crucial for monitoring disease progression and evaluating therapeutic response. EXPERT OPINION: Growing evidence supports HSCT as a disease-modifying strategy that can stabilize or slow progression, particularly when performed early in the symptomatic phase. However, HSCT's invasiveness and associated risks highlight the need for safer, more targeted interventions. The future of CSF1R-RD treatment lies in developing less invasive microglia replacement methods and enhancing engraftment efficacy, leveraging newly established clinical and biomarker tools for precise monitoring.

  PublisherDOI

• Human microglia differentially respond to β‐amyloid, tau, and combined Alzheimer's disease pathologies in vivo

  Alzheimer s & Dementia · 2025-11-01 · 5 citations

  articleOpen access

  INTRODUCTION: Recent studies have identified important species-dependent differences in the response of microglia to β-amyloid (Aβ) pathology. Yet, whether human microglia also interact differently with the pathognomonic combination of amyloid and tau pathologies that occur in Alzheimer's disease (AD) remains unclear. METHODS: We generated a xenotolerant mouse model of AD that develops both plaque and tangle pathologies, transplanted stem cell-derived microglial progenitors and examined the interactions between human microglia and AD pathologies with scRNA sequencing, immunohistochemistry, and in vitro modeling. RESULTS: The combined amyloid and tau pathologies induced robust type-I interferon and proinflammatory cytokine responses, as well as an increased adoption of a distinct "rod" morphology in human microglia. The rod morphology could be induced with type-I interferon treatment in vitro. DISCUSSION: We provide new insights into human microglial responses to combined AD pathologies and a novel platform to investigate and manipulate human microglia in vivo. HIGHLIGHTS: Amyloid pathology promotes the rapid development of neurofibrillary tangles and neuronal loss in a novel chimeric model of AD. Combined Alzheimer's disease pathologies lead to an expansion of disease-associated microglia (DAM) and exacerbate Interferon-responsive and cytokine/chemokine-enriched states in xenotransplanted human microglia. The combination of amyloid and tau promotes the development of a distinctive rod microglial phenotype that closely correlates with tau pathology and neurodegeneration. Rod morphology and transcriptional changes can be modeled in vitro by treatment of induced pluripotent stem cells (iPSC) -microglia with type-I interferons.

  PublisherDOI

• Fluid and Neuroimaging Biomarkers in Microgliopathy Colony-Stimulating Factor-1 Receptor-Related Disorders

  medRxiv · 2025-09-14

  preprintOpen access

  Abstract Colony-stimulating factor 1 receptor-related disorder (CSF1R-RD) is a neurodegenerative condition characterized by rapid progression, leading to profound functional decline and ultimately resulting in a persistent vegetative state. Although an effective treatment option exists, there remains a lack of identified biomarkers capable of monitoring disease progression and detecting the earliest symptom onset in CSF1R pathogenic variant carriers, limiting the ability of clinicians to make informed decisions regarding patient care. This study aims to identify both fluid and neuroimaging biomarkers for CSF1R-RD that can inform the optimal timing of treatment administration to maximize therapeutic benefit, while also providing sensitive quantitative measurements to monitor disease progression. Our study compared neuroimaging and fluid (plasma and cerebrospinal fluid (CSF)) biomarkers across three distinct populations: asymptomatic CSF1R pathogenic variant carriers (N=14), symptomatic CSF1R pathogenic variant carriers (N=17), and healthy controls (N=30). We evaluated biomarker correlations with both an established (Montreal Cognitive Assessment (MoCA)) and a novel (CSF1R Clinical Severity Score (CCSS)) clinical diagnostic scale to investigate potential clinical utility. Additionally, we tested the relationship between select biomarkers and cortical thickness using 3D T1-weighted MPRAGE scans, providing a highly valuable physiological component to our analyses. Our results demonstrate that while plasma glial fibrillary acidic protein (GFAP) displays a high sensitivity for distinguishing early-stage CSF1R-RD patients from healthy controls, plasma neurofilament light chain (NfL) is more effective for tracking disease progression following the onset of symptoms. Overall, our study provides evidence for plasma NfL and GFAP as valuable biomarkers of earliest symptom onset and disease progression for CSF1R-RD One Sentence Summary This study identifies plasma biomarkers NfL and GFAP as promising tools to detect CSF1R-RD onset and progression, with potential to improve patient outcomes.

  PublisherOA PDFDOI

• Harnessing human iPSC-microglia for CNS-wide delivery of disease-modifying proteins

  Cell stem cell · 2025-04-14 · 39 citations

  articleOpen access

  Widespread delivery of therapeutic proteins to the brain remains challenging. To determine whether human induced pluripotent stem cell (iPSC)-microglia (iMG) could enable brain-wide and pathology-responsive delivery of therapeutic cargo, we utilized CRISPR gene editing to engineer iMG to express the Aβ-degrading enzyme neprilysin under control of the plaque-responsive promoter, CD9. To further determine whether increased engraftment enhances efficacy, we utilized a CSF1R-inhibitor resistance approach. Interestingly, both localized and brain-wide engraftment in Alzheimer's disease (AD) mice reduced multiple biochemical measures of pathology. However, within the plaque-dense subiculum, reductions in plaque load, dystrophic neurites, and astrogliosis and preservation of neuronal density were only achieved following widespread microglial engraftment. Lastly, we examined chimeric models of breast cancer brain metastases and demyelination, demonstrating that iMG adopt diverse transcriptional responses to differing neuropathologies, which could be harnessed to enable widespread and pathology-responsive delivery of therapeutics to the CNS.

  PublisherDOI

• Novel photocrosslinking chemical probes utilized for high-resolution spatial transcriptomics

  RSC Chemical Biology · 2025-01-01 · 1 citations

  articleOpen accessSenior authorCorresponding

  the exterior population. We anticipate this new technology will be a valuable tool addition as it will enable easier access to spatial transcriptomic analysis for the scientific community using conventional microscopy and analysis techniques.

  PublisherOA PDFDOI

• Aberrant splicing in Huntington’s disease accompanies disrupted TDP-43 activity and altered m6A RNA modification

  Nature Neuroscience · 2025-01-06 · 33 citations

  articleOpen accessCorresponding

  Huntington's disease (HD) is caused by a CAG repeat expansion in the HTT gene, leading to altered gene expression. However, the mechanisms leading to disrupted RNA processing in HD remain unclear. Here we identify TDP-43 and the N6-methyladenosine (m6A) writer protein METTL3 to be upstream regulators of exon skipping in multiple HD systems. Disrupted nuclear localization of TDP-43 and cytoplasmic accumulation of phosphorylated TDP-43 occurs in HD mouse and human brains, with TDP-43 also co-localizing with HTT nuclear aggregate-like bodies distinct from mutant HTT inclusions. The binding of TDP-43 onto RNAs encoding HD-associated differentially expressed and aberrantly spliced genes is decreased. Finally, m6A RNA modification is reduced on RNAs abnormally expressed in the striatum of HD R6/2 mouse brain, including at clustered sites adjacent to TDP-43 binding sites. Our evidence supports TDP-43 loss of function coupled with altered m6A modification as a mechanism underlying alternative splicing in HD.

  PublisherOA PDFDOI

• Quantification of subcellular RNA localization through direct detection of RNA oxidation

  Nucleic Acids Research · 2025-02-12 · 10 citations

  articleOpen access

  Across cell types and organisms, thousands of RNAs display asymmetric subcellular distributions. Studying this process requires quantifying abundances of specific RNAs at precise subcellular locations. To analyze subcellular transcriptomes, multiple proximity-based techniques have been developed in which RNAs near a localized bait protein are specifically labeled, facilitating their biotinylation and purification. However, these complex methods are often laborious and require expensive enrichment reagents. To streamline the analysis of localized RNA populations, we developed Oxidation-Induced Nucleotide Conversion sequencing (OINC-seq). In OINC-seq, RNAs near a genetically encoded, localized bait protein are specifically oxidized in a photo-controllable manner. These oxidation events are then directly detected and quantified using high-throughput sequencing and our software package, PIGPEN, without the need for biotin-mediated enrichment. We demonstrate that OINC-seq can induce and quantify RNA oxidation with high specificity in a dose- and light-dependent manner. We further show the spatial specificity of OINC-seq by using it to quantify subcellular transcriptomes associated with the cytoplasm, ER, nucleus, and the inner and outer membranes of mitochondria. Finally, using transgenic zebrafish, we demonstrate that OINC-seq allows proximity-mediated RNA labeling in live animals. In sum, OINC-seq together with PIGPEN provide an accessible workflow for analyzing localized RNAs across different biological systems.

  PublisherDOI

• Therapeutic potential of human microglia transplantation in a chimeric model of CSF1R-related leukoencephalopathy

  Neuron · 2024-06-18 · 76 citations

  articleOpen access

  Microglia replacement strategies are increasingly being considered for the treatment of primary microgliopathies like adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). However, available mouse models fail to recapitulate the diverse neuropathologies and reduced microglia numbers observed in patients. In this study, we generated a xenotolerant mouse model lacking the fms-intronic regulatory element (FIRE) enhancer within Csf1r, which develops nearly all the hallmark pathologies associated with ALSP. Remarkably, transplantation of human induced pluripotent stem cell (iPSC)-derived microglial (iMG) progenitors restores a homeostatic microglial signature and prevents the development of axonal spheroids, white matter abnormalities, reactive astrocytosis, and brain calcifications. Furthermore, transplantation of CRISPR-corrected ALSP-patient-derived iMG reverses pre-existing spheroids, astrogliosis, and calcification pathologies. Together with the accompanying study by Munro and colleagues, our results demonstrate the utility of FIRE mice to model ALSP and provide compelling evidence that iMG transplantation could offer a promising new therapeutic strategy for ALSP and perhaps other microglia-associated neurological disorders.

  PublisherOA PDFDOI

Recent grants

• A novel platform for the investigation of human microglia

  NIH · $2.1M · 2021–2023

• Determining the factors that control dose-dependent splicing regulation by a master regulator

  NIH · $2.0M · 2017–2022

• Multicellular simultaneous chemotranscriptomic profiling of nascent transcription during learning

  NIH · $425k · 2017–2020

• Cracking the RNA Localization Code

  NIH · $2.2M · 2015–2020

• Epitranscriptomic mechanisms of fear-related learning and memory

  NIH · $1.9M · 2016–2022

Frequent coauthors

• Howard Y. Chang

  Stanford University

  37 shared

• Whitney England

  University of California, Irvine

  27 shared

• Ryan A. Flynn

  25 shared

• Chao Feng

  People's Liberation Army 401 Hospital

  25 shared

• Joseph E. Wedekind

  University of Rochester

  24 shared

• Timothy W. Bredy

  University of California, Irvine

  21 shared

• J. Krucinska

  University of Connecticut

  21 shared

• Gloria Cristalli

  Università di Camerino

  18 shared

Education

• Ph.D., Chemistry

  University of Rochester

  2009