Science

Coming Together in Times of Crisis
Cell and molecular biology of Regulated IRE1α-Mediated Decay (RIDD) in epithelial fate transitions
Getting Back on Track
Fate and function of epithelial transitional states in pulmonary fibrosis
Stepping Back From the Brink
Regulation of goblet cell metaplasia in asthma and other muco-obstructive diseases

Projects

Cell and molecular biology of Regulated IRE1α-Mediated Decay (RIDD) in epithelial fate transitions

IRE1α is an endoplasmic reticulum (ER) transmembrane protein containing a luminal domain that is responsive to protein-folding stress (ER stress). IRE1α is ancient and its functions are conserved throughout eukaryotes. Upon stress, IRE1α kinase domains self-associate and trans-autophosphorylate, which activates the RNase domain to cleave the mRNA of the XBP1 transcription factor and initiate its alternative splicing. Spliced XBP1 upregulates factors that promote ER protein folding including chaperones and oxidoreductases. Under severe ER stress, IRE1α degrades a plethora of mRNAs localizing to the ER membrane and select micro-RNAs in a process dubbed Regulated IRE1α-Dependent Decay (RIDD).

Though protein folding homeostasis was IRE1α's ancestral function, we think it evolved to take on more complex roles as multicellular organisms developed. In particular, our work on epithlelial transition states in fibrosis suggests that RIDD regulates epithelial plasticity by degrading mRNAs encoding genes important to maintaining cellular identity. These genes include some that are not predicted to be translated at the ER membrane. We are exploring the idea that enzyme and substrate only come together during crisis: that cell state and identity causes recruitment of select mRNAs to IRE1α on the cytosolic face of the ER.

Fate and function of epithelial transitional states in pulmonary fibrosis

Idiopathic pulmononary fibrosis (IPF), the most common form of lung fibrosis, is a disease of progressive respiratory failure with limited treatment options.  A new paradigm recently emerged in the biology of lung fibrosis: in the course of repairing the lung, facultative stem cells enter "transitional" states that can actually be pathological. On one hand, transitional states can successfully differentiate into newly regenerated lung tissue, but on the other hand, failure to differentiate can instead nucleate fibrotic disease.

Using single-cell sequencing, we have found that cells enter a particular transitional state that is uniquely found in models of fibrosis and human fibrotic lungs. We hypothesize that fibrotic lung injury causes lung epithelial stem cells to detour into a dead-end pathologic state, where they are unable to differentiate into new alveolar epithelial cells. We are using new genetic tracing tools to follow the fates of cells in the fibrotic state, and new spatial transcriptomic platforms to characterize their interactions with other cells in the fibrotic niche.

The niche signals and cell-intrinsic pathways that drive cells into the fibrotic dead-end state are incompletely understood. Cells in the state exhibit features of senescence and cellular stress, including a signature of unfolded protein response (UPR) and IRE1α activation. Using genetic tools and unique small-molecule modulators of the IRE1α kinase, we have shown that inhibiting IRE1α reduces fibrosis and promotes successful differentiation. We using a new generation of IRE1α modulators to understand the molecular mechanisms of how IRE1α regulates the fibrotic state. Our goal is to use small molecules to modulate IRE1α and other signals/pathways and get cells back onto a regenerative track to repair the lung.

Regulation of goblet cell metaplasia in asthma and other muco-obstructive diseases

Mucus hypersecretion causes symptoms and reduces lung function in human asthma and other lung diseases, including COPD and chronic rhinosinusitis. Despite advances in targeted therapies to reduce inflammation in asthma, there are no therapies that directly target mucus hypersecretion.

In human airways, basal cells differentiate into mucin-producing goblet cells in response to allergic inflammatory signals. In collaboration with the UCSF Asthma and Allergic Diseases Cooperative Research Center, we have found that the unfolded protein response (UPR) and IRE1α are activated during differentiation. Using genetic tools and small-molecule modulators of the IRE1α kinase, we have shown that inhibiting IRE1α reduces goblet cell metaplasia in vivo and in air-liquid interface culture. We using a new generation of IRE1α modulators to understand the molecular mechanisms of how IRE1α regulates goblet cell maturation. Our goal is to use small molecules to inhibit IRE1α and get airway epithelial cells to "step back from the brink" of the goblet cell fate, and thus reduce mucus and symptom burden in asthma.


Publications

Highlights

Auyeung VC, Downey MS, Thamsen M, Wenger TA, Backes BJ, Sheppard D, Papa FR. IRE1α drives lung epithelial progenitor dysfunction to establish a niche for pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol. 2022 Apr 1;322(4):L564-L580. doi: 10.1152/ajplung.00408.2021. Epub 2022 Feb 16. PubMed PMID: 35170357; PubMed Central PMCID: PMC8957349.

Feldman HC, Ghosh R, Auyeung VC, Mueller JL, Kim JH, Potter ZE, Vidadala VN, Perera BGK, Olivier A, Backes BJ, Zikherman J, Papa FR, Maly DJ. ATP-competitive partial antagonists of the IRE1α RNase segregate outputs of the UPR. Nat Chem Biol. 2021 Nov;17(11):1148-1156. doi: 10.1038/s41589-021-00852-0. Epub 2021 Sep 23. PubMed PMID: 34556859; PubMed Central PMCID: PMC8551014.

Chronological

Le Saux CJ, Ho TC, Brumwell AM, Kathiriya JJ, Wei Y, Hughes JB, Garakani K, Atabai K, Auyeung VC, Papa FR, Chapman HA. BCL-2 Modulates IRE1α Activation to Attenuate Endoplasmic Reticulum Stress and Pulmonary Fibrosis. Am J Respir Cell Mol Biol. 2024 Apr;70(4):247-258. doi: 10.1165/rcmb.2023-0109OC. PubMed PMID: 38117250.
 
Auyeung VC, Downey MS, Thamsen M, Wenger TA, Backes BJ, Sheppard D, Papa FR. IRE1α drives lung epithelial progenitor dysfunction to establish a niche for pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol. 2022 Apr 1;322(4):L564-L580. doi: 10.1152/ajplung.00408.2021. Epub 2022 Feb 16. PubMed PMID: 35170357; PubMed Central PMCID: PMC8957349.
 
Feldman HC, Ghosh R, Auyeung VC, Mueller JL, Kim JH, Potter ZE, Vidadala VN, Perera BGK, Olivier A, Backes BJ, Zikherman J, Papa FR, Maly DJ. ATP-competitive partial antagonists of the IRE1α RNase segregate outputs of the UPR. Nat Chem Biol. 2021 Nov;17(11):1148-1156. doi: 10.1038/s41589-021-00852-0. Epub 2021 Sep 23. PubMed PMID: 34556859; PubMed Central PMCID: PMC8551014.
 
Auyeung VC, Sheppard D. Stuck in a Moment: Does Abnormal Persistence of Epithelial Progenitors Drive Pulmonary Fibrosis?. Am J Respir Crit Care Med. 2021 Mar 15;203(6):667-669. doi: 10.1164/rccm.202010-3898ED. PubMed PMID: 33226835; PubMed Central PMCID: PMC7958518.
 
Gonzalez-Teuber V, Albert-Gasco H, Auyeung VC, Papa FR, Mallucci GR, Hetz C. Small Molecules to Improve ER Proteostasis in Disease. Trends Pharmacol Sci. 2019 Sep;40(9):684-695. doi: 10.1016/j.tips.2019.07.003. Epub 2019 Jul 31. Review. PubMed PMID: 31377018.
 
Thamsen M, Ghosh R, Auyeung VC, Brumwell A, Chapman HA, Backes BJ, Perara G, Maly DJ, Sheppard D, Papa FR. Small molecule inhibition of IRE1α kinase/RNase has anti-fibrotic effects in the lung. PLoS One. 2019;14(1):e0209824. doi: 10.1371/journal.pone.0209824. eCollection 2019. PubMed PMID: 30625178; PubMed Central PMCID: PMC6326459.
 
Auyeung VC, Ulitsky I, McGeary SE, Bartel DP. Beyond secondary structure: primary-sequence determinants license pri-miRNA hairpins for processing. Cell. 2013 Feb 14;152(4):844-58. doi: 10.1016/j.cell.2013.01.031. PubMed PMID: 23415231; PubMed Central PMCID: PMC3707628.
 
Chiang HR, Schoenfeld LW, Ruby JG, Auyeung VC, Spies N, Baek D, Johnston WK, Russ C, Luo S, Babiarz JE, Blelloch R, Schroth GP, Nusbaum C, Bartel DP. Mammalian microRNAs: experimental evaluation of novel and previously annotated genes. Genes Dev. 2010 May 15;24(10):992-1009. doi: 10.1101/gad.1884710. Epub 2010 Apr 22. PubMed PMID: 20413612; PubMed Central PMCID: PMC2867214.
 
D'Alise AM, Auyeung V, Feuerer M, Nishio J, Fontenot J, Benoist C, Mathis D. The defect in T-cell regulation in NOD mice is an effect on the T-cell effectors. Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19857-62. doi: 10.1073/pnas.0810713105. Epub 2008 Dec 10. PubMed PMID: 19073938; PubMed Central PMCID: PMC2604930.
 
An DS, Qin FX, Auyeung VC, Mao SH, Kung SK, Baltimore D, Chen IS. Optimization and functional effects of stable short hairpin RNA expression in primary human lymphocytes via lentiviral vectors. Mol Ther. 2006 Oct;14(4):494-504. doi: 10.1016/j.ymthe.2006.05.015. Epub 2006 Jul 17. PubMed PMID: 16844419; PubMed Central PMCID: PMC2562632.
 
Dahan DS, Dibas MI, Petersson EJ, Auyeung VC, Chanda B, Bezanilla F, Dougherty DA, Lester HA. A fluorophore attached to nicotinic acetylcholine receptor beta M2 detects productive binding of agonist to the alpha delta site. Proc Natl Acad Sci U S A. 2004 Jul 6;101(27):10195-200. doi: 10.1073/pnas.0301885101. Epub 2004 Jun 24. PubMed PMID: 15218096; PubMed Central PMCID: PMC454187.