Biology subjects

Schaeffer, A. J.

Publications and source records attributed to Schaeffer, A. J..

6 recordsLinked to original sources

bioRxiv2026

MULTI-COHORT TRANSCRIPTOMIC ANALYSIS REVEALS SHARED INFLAMMATORY SIGNALING AND UROTHELIAL DYSFUNCTION ACROSS IC/BPS PHENOTYPES

Interstitial cystitis/bladder pain syndrome (IC/BPS) is a heterogeneous disorder characterized by bladder pain, inflammation, and urothelial dysfunction, yet the molecular features shared across clinically defined disease phenotypes remain incompletely understood. Toll-like receptor 4 (TLR4) and acyloxyacyl hydrolase (AOAH), a regulator of TLR4 ligand activity, have been implicated in IC/BPS in clinical and preclinical mechanistic studies. Here, we performed a multi-cohort reanalysis of four independent human IC/BPS transcriptomic datasets spanning bladder tissue and urine sediment from non-ulcer/non-Hunner, ulcerative/Hunner, normal-capacity, low-capacity, non-lesion, and lesion phenotypes. We evaluated immune cell-associated expression, urothelial differentiation and barrier integrity, TLR4 and pro-inflammatory signaling, and AOAH expression. IC/BPS exhibited phenotype-specific immune remodeling, yet shared features were also identified. Bladder-tissue IC/BPS was consistently associated with reduced expression of urothelial differentiation and barrier markers, together with increased TLR4 recognition and pro-inflammatory signaling. AOAH expression was elevated across all three bladder-tissue cohorts and was associated with monocyte/macrophage and pro-inflammatory transcriptional states. TLR4 signaling similarly paralleled myeloid and inflammatory features, whereas inflammatory programs generally showed inverse relationships with urothelial barrier markers. Notably, independently defined severe phenotypes of ulcerative IC and low-capacity IC showed concordant molecular remodeling characterized by increased leukocyte and monocyte/macrophage signatures, enhanced TLR4 signaling, elevated AOAH expression, and reduced urothelial barrier-associated expression. Together, these findings identify reproducible molecular signatures that extend across IC/BPS phenotypes and support a model in which urothelial dysfunction and TLR4-AOAH-associated myeloid inflammatory states coexist across disease phenotypes. These results provide a molecular framework for stratifying IC/BPS beyond clinical labels and identify shared and subtype-specific molecular signatures that may inform therapeutic response evaluation and clinical management.

immunology↗

bioRxiv2026

PD-1-Dependent Modulation of Sensory Neurons by a Staphylococcus epidermidis Lipoteichoic Acid Drives Analgesia

BackgroundImmune checkpoint receptors regulate adaptive immunity but are increasingly recognized as modulators of neuroimmune interactions. The upstream signals that induce neuronal checkpoint pathways during inflammation and their functional relevance in inflammatory pain remain incompletely understood. We investigated whether a defined commensal-derived molecule engages a neuroimmune checkpoint axis to modulate inflammatory pain. MethodsSELTA, a lipoteichoic acid purified from a commensal Staphylococcus epidermidis strain, was evaluated for TLR2-dependent activity, regulation of Pdcd1 transcription and PD-1 protein expression in dorsal root ganglion (DRG) neurons, effects on intracellular calcium signaling, and behavioral outcomes in experimental autoimmune prostatitis (EAP) a model of inflammation-induced chronic pelvic pain. Conditional Pdcd1 deletion was performed in sensory neurons (Advillin-Cre) and CD4 T cells to determine cell-specific requirements. ResultsSELTA selectively activated TLR2/6 signaling and increased Pdcd1 transcription and PD-1 protein expression in DRG neurons under inflammatory conditions. SELTA enhanced phosphorylation of PD-1 at tyrosine 248 and significantly reduced ATP-evoked intracellular Ca{superscript 2} responses in mouse primary sensory neurons. Pharmacologic neutralization of PD-1 abrogated SELTA-mediated suppression of calcium signaling. In vivo, SELTA produced concentration-dependent attenuation of pelvic hypersensitivity in EAP. Conditional deletion of PD-1 in vivo in Advillin-expressing sensory neurons or CD4 T cells significantly reduced SELTA-induced analgesia, while combined deletion did not further diminish the effect. ConclusionsThese findings identify a commensal-derived lipoteichoic acid that engages a TLR2/6-associated pathway linked to inducible neuronal PD-1 signaling to restrain inflammatory nociceptor activity. The results define a neuroimmune checkpoint mechanism that modulates inflammatory pain and extend PD-1 signaling beyond its established role in adaptive immunity.

neuroscience↗

bioRxiv2021

Mast cell function in prostate inflammation, fibrosis, and smooth muscle cell dysfunction

Intraurethral inoculation of mice with uropathogenic E. coli (CP1) results in prostate inflammation, fibrosis, and urinary dysfunction, recapitulating some but not all of the pathognomonic clinical features associated with benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS). In both patients with LUTS and in CP1-infected mice, we observed increased numbers and activation of mast cells and elevated levels of prostate fibrosis. Therapeutic inhibition of mast cells and the histamine 1 receptor in the mouse model resulted in reduced mast cell activation in the prostate and significant alleviation of urinary dysfunction. Treated mice showed reduced prostate fibrosis, less infiltration of immune cells, and decreased inflammation. In addition, as opposed to symptomatic CP1-infected mice, treated mice showed reduced myosin light chain (MLC)-2 phosphorylation, a marker of prostate smooth muscle contraction. These results show that mast cells play a critical role in the pathophysiology of urinary dysfunction and may be an important therapeutic target for men with BPH/LUTS. NEW AND NOTEWORTHYLUTS-associated BPH is derived from a combination of immune activation, extracellular matrix remodeling, hyperplasia, and smooth muscle cell contraction in prostates of men. Using a mouse model, we describe the importance of mast cells in regulating these multiple facets involved in the pathophysiology of LUTS. Mast cell inhibition alleviates both pathology and urinary dysfunction in this model suggesting the potential for mast cell inhibition as a therapeutic that prevents and reverses pathology and associated symptomology.

cell biology↗

bioRxiv2021

Acyloxyacyl Hydrolase Modulates the Gut Microbiome Through Transcriptional Regulators of Corticotropin-Releasing Factor

Gut microbiome-host interactions play a crucial role in health and disease. Altered gut microbiome composition has been observed in patients with interstitial cystitis/bladder pain syndrome (IC/BPS), a disorder characterized by pelvic pain, voiding dysfunction, and often co-morbid with anxiety/depression. We recently showed that mice deficient for acyloxyacyl hydrolase (AOAH) mimic pelvic pain symptoms and comorbidities of IC/BPS and also exhibit gut dysbiosis. In addition, we previously identified that the conditional knockout (cKO) of two transcriptional regulators of the gene encoding corticotropin-releasing factor, Crf, that are downstream of AOAH, aryl hydrocarbon receptor (AhR) and peroxisome proliferator-activated receptor-{gamma} (PPAR{gamma}), alleviate anxiety/depressive and voiding phenotypes of AOAH-deficient mice. Here, we examined the effects of AhR and PPAR{gamma} in CRF-expressing cells on the dysbiosis of AOAH-deficiency. AOAH-deficient mice with cKO of PPAR{gamma} and AhR/PPAR{gamma} exhibited reduced pelvic allodynia compared to AOAH-deficient mice, suggesting a role for PPAR{gamma} in regulating pelvic pain. 16S rRNA sequencing of fecal stool from female AOAH-deficient mice with a cKO of AhR and/or PPAR{gamma} in CRF-expressing cells identified altered gut microbiota distinct from AOAH-deficient stool. The cKO of AhR and PPAR{gamma} showed improved cecum barrier function in females compared to AOAH-deficient mice, whereas males were primarily affected by PPAR{gamma}, suggesting sex differences in gut responses. Pair-wise comparison of microbiota also suggested sex differences in response to AOAH-deficiency and conditional knockout of AhR and PPAR{gamma}. Our findings suggest that the dysbiosis and leaky gut of AOAH deficiency is mediated by AhR and PPAR{gamma} in CRF-expressing cells and reveal a novel mechanism and therapeutic targets for pelvic pain.

systems biology↗

bioRxiv2021

Acyloxyacyl Hydrolase Regulates Microglia-Mediated Pelvic Pain Through Toll-Like Receptor-4

Interstitial cystitis/bladder pain syndrome (IC/BPS) is a devastating condition of chronic pelvic pain and urinary dysfunction. We have shown that mice deficient for the lipase acyloxyacyl hydrolase (AOAH) develop pelvic allodynia and exhibit symptoms and comorbidities consistent with IC/BPS, as well as gut dysbiosis. Microglia are resident immune cells of the central nervous system (CNS) that respond to changes in the gut microbiome, and studies have linked microglial activation to neuropathic pain. Additionally, microglia express toll-like receptors (TLRs), including TLR4, which are activated by microbial components. We have previously shown that AOAH-deficient mice exhibit increased gut permeability, suggesting a possible mechanism of microglial TLR4 activation via translocation of microbial products across the intestinal barrier to the brain. Here, we assessed the role of AOAH and TLR4 in microglial activation and pelvic pain. AOAH immunoreactivity co-localized with the microglial marker P2YR12 but not astrocytes, suggesting a functional role for AOAH in microglia. Pharmacologic ablation of CNS microglia with PLX5622 resulted in decreased pelvic allodynia in AOAH-deficient mice and resurgence of pelvic pain upon drug washout. Aligned with microglial activation, we observed altered cytokine abundance in Aoah-/- cortex that was reduced in Aoah/Tlr4-/- cortex. Consistent with our hypothesis of TLR4 activation by gut microbes, we observed microbiome-dependent activation of cultured BV2 microglial cells. Skeletal analyses revealed that AOAH-deficient mice have an activated microglia morphology in brain regions associated with neuropathic pain, independent of TLR4. Compared to Aoah-/- mice, Aoah/Tlr4-/- mice exhibited decreased pelvic pain and microglial cytokine expression. Together, these findings demonstrate differential roles for AOAH and TLR4 in microglial activation and pelvic pain and thus identify novel therapeutic targets for IC/BPS.

neuroscience↗

bioRxiv2021

Acyloxyacyl Hydrolase is a Host Determinant of Gut Microbiome-Mediated Pelvic Pain

Dysbiosis of gut microbiota is associated with many pathologies, yet host factors modulating microbiota remain unclear. Interstitial cystitis/bladder pain syndrome (IC/BPS or "IC") is a debilitating condition of chronic pelvic pain often with co-morbid urinary dysfunction and anxiety/depression, and recent studies find fecal dysbiosis in IC/BPS patients. We previously identified the locus encoding acyloxyacyl hydrolase, Aoah, as a modulator of pelvic pain severity in a murine IC/BPS model. AOAH-deficient mice spontaneously develop rodent correlates of pelvic pain, increased responses to induced pelvic pain models, voiding dysfunction, and anxious/depressive behaviors. Here, we report that AOAH-deficient mice exhibit dysbiosis of GI microbiota. AOAH-deficient mice exhibit an enlarged cecum, a phenotype long associated with germ-free rodents, and reduced trans-epithelial electrical resistance consistent with a "leaky gut" phenotype. AOAH-deficient ceca showed altered gene expression consistent with inflammation, Wnt signaling, and urologic disease. 16S rRNA sequencing of stool revealed altered microbiota in AOAH-deficient mice, and GC-MS identified altered metabolomes. Co-housing AOAH-deficient mice with wild type mice resulted in converged microbiota and altered predicted metagenomes. Co-housing also abrogated the pelvic pain phenotype of AOAH-deficient mice, which was corroborated by oral gavage of AOAH-deficient mice with stool slurry of wild type mice. Converged microbiota also alleviated comorbid anxiety-like behavior in AOAH-deficient mice. Oral gavage of AOAH-deficient mice with anaerobes cultured from IC/BPS stool resulted in exacerbation of pelvic allodynia. Together, these data indicate that AOAH is a host determinant of normal gut microbiota, and the dysbiosis associated with AOAH deficiency contributes to pelvic pain. These findings suggest that the gut microbiome is a potential therapeutic target for IC/BPS.

systems biology↗