TNFR1 membrane reorganization promotes distinct modes of TNFα signaling.

Signaling by the ubiquitously expressed tumor necrosis factor receptor 1 (TNFR1) after ligand binding plays an essential role in determining whether cells exhibit survival or death. TNFR1 forms distinct signaling complexes that initiate gene expression programs downstream of the transcriptional regulators NFκB and AP-1 and promote different functional outcomes, such as inflammation, apoptosis, and necroptosis. Here, we investigated the ways in which TNFR1 was organized at the plasma membrane at the nanoscale level to elicit different signaling outcomes. We confirmed that TNFR1 forms preassembled clusters at the plasma membrane of adherent cells in the absence of ligand. After trimeric TNFα binding, TNFR1 clusters underwent a conformational change, which promoted lateral mobility, their association with the kinase MEKK1, and activation of the JNK/p38/NFκB pathway. These phenotypes required a minimum of two TNFR1-TNFα contact sites; fewer binding sites resulted in activation of NFκB but not JNK and p38. These data suggest that distinct modes of TNFR1 signaling depend on nanoscale changes in receptor organization.

OSSOS XV: PROBING THE DISTANT SOLAR SYSTEM WITH OBSERVED SCATTERING TNOS.

Most known trans-Neptunian objects (TNOs) gravitationally scattering off the giant planets have orbital inclinations consistent with an origin from the classical Kuiper belt, but a small fraction of these "scattering TNOs" have inclinations that are far too large (i > 45°) for this origin. These scattering outliers have previously been proposed to be interlopers from the Oort cloud or evidence of an undiscovered planet. Here we test these hypotheses using N-body simulations and the 69 centaurs and scattering TNOs detected in the Outer Solar Systems Origins Survey and its predecessors. We confirm that observed scattering objects cannot solely originate from the classical Kuiper belt, and we show that both the Oort cloud and a distant planet generate observable highly inclined scatterers. Although the number of highly inclined scatterers from the Oort Cloud is ~3 times less than observed, Oort cloud enrichment from the Sun's galactic migration or birth cluster could resolve this. Meanwhile, a distant, low-eccentricity 5 M⊕ planet replicates the observed fraction of highly inclined scatterers, but the overall inclination distribution is more excited than observed. Furthermore, the distant planet generates a longitudinal asymmetry among detached TNOs that is less extreme than often presumed, and its direction reverses across the perihelion range spanned by known TNOs. More complete models that explore the dynamical origins of the planet are necessary to further study these features. With observational biases well-characterized, our work shows that the orbital distribution of detected scattering bodies is a powerful constraint on the unobserved distant solar system.

KIF22 coordinates CAR and EGFR dynamics to promote cancer cell proliferation.

The coxsackievirus and adenovirus receptor (CAR) is a transmembrane receptor that plays a key role in cell-cell adhesion. CAR is found in normal epithelial cells and is increased in abundance in various human tumors, including lung carcinomas. We investigated the potential mechanisms by which CAR contributes to cancer cell growth and found that depletion of CAR in human lung cancer cells reduced anchorage-independent growth, epidermal growth factor (EGF)-dependent proliferation, and tumor growth in vivo. EGF induced the phosphorylation of CAR and its subsequent relocalization to cell junctions through the activation of the kinase PKCδ. EGF promoted the binding of CAR to the chromokinesin KIF22. KIF22-dependent regulation of microtubule dynamics led to delayed EGFR internalization, enhanced EGFR signaling, and coordination of CAR dynamics at cell-cell junctions. These data suggest a role for KIF22 in the coordination of membrane receptors and provide potential new therapeutic strategies to combat lung tumor growth.

TNFα promotes CAR-dependent migration of leukocytes across epithelial monolayers.

Trans-epithelial migration (TEpM) of leukocytes during inflammation requires engagement with receptors expressed on the basolateral surface of the epithelium. One such receptor is Coxsackie and Adenovirus Receptor (CAR) that binds to Junctional Adhesion Molecule-like (JAM-L) expressed on leukocytes. Here we provide the first evidence that efficient TEpM of monocyte-derived THP-1 cells requires and is controlled by phosphorylation of CAR. We show that TNFα acts in a paracrine manner on epithelial cells via a TNFR1-PI3K-PKCδ pathway leading to CAR phosphorylation and subsequent transmigration across cell junctions. Moreover, we show that CAR is hyper-phosphorylated in vivo in acute and chronic lung inflammation models and this response is required to facilitate immune cell recruitment. This represents a novel mechanism of feedback between leukocytes and epithelial cells during TEpM and may be important in controlling responses to pro-inflammatory cytokines in pathological settings.