bims-tucedo 2022-01-30 papers

bims-tucedo

Biomed News

on Tumor cell dormancy

Issue of 2022–01–30
twenty papers selected by
Isabel Puig Borreil, Vall d’Hebron Institute of Oncology

Senescence induction dictates response to chemo- and immunotherapy in preclinical models of ovarian cancer.
Hotspot ESR1 mutations are multimodal and contextual modulators of breast cancer metastasis.
Mapping molecular subtype specific alterations in breast cancer brain metastases identifies clinically relevant vulnerabilities.
Targeting S100A9-ALDH1A1-retinoic acid signaling to suppress brain relapse in EGFR-mutant lung cancer.
Smad3 methylation by EZH2 promotes its activation and tumor metastasis.
IKBKE phosphorylates and stabilizes Snail to promote breast cancer invasion and metastasis.
Functional analysis of MET exon 14 skipping alteration in cancer invasion and metastatic dissemination.
Medical treatment for active breast cancer brain metastases.
Tumor ENPP1(CD203a)/Haptoglobin Axis Exploits Myeloid-Derived Suppressor Cells to Promote Post-Radiotherapy Local Recurrence in Breast Cancer.
Endothelial p130cas confers resistance to anti-angiogenesis therapy.
The circROBO1/KLF5/FUS feedback loop regulates the liver metastasis of breast cancer by inhibiting the selective autophagy of afadin.
Targeting a splicing-mediated drug resistance mechanism in prostate cancer by inhibiting transcriptional regulation by PKCβ1.
Loss of miR-31-5p drives hematopoietic stem cell malignant transformation and restoration eliminates leukemia stem cells in mice.
Tumour-associated neutrophils secrete AGR2 to promote colorectal cancer metastasis via its receptor CD98hc-xCT.
Canonical TGFβ signaling induces collective invasion in colorectal carcinogenesis through a Snail1- and Zeb1-independent partial EMT.
Disruption of pancreatic stellate cell myofibroblast phenotype promotes pancreatic tumor invasion.
Single cell DNA-seq depicts clonal evolution of multiple driver alterations in osimertinib resistant patients.
Opioid receptor signaling suppresses leukemia through both catalytic and non-catalytic functions of TET2.
Quality and Quantity: Evaluating Tumor Biology Alongside Novel Imaging on Diagnosis of Metastatic Hormone-sensitive Prostate Cancer.
FREQUENCY AND GENOMIC ASPECTS OF INTRINSIC RESISTANCE TO VISMODEGIB IN LOCALLY ADVANCED BASAL CELL CARCINOMA.

Proc Natl Acad Sci U S A. 2022 Feb 01. pii: e2117754119. [Epub ahead of print]119(5):

Senescence induction dictates response to chemo- and immunotherapy in preclinical models of ovarian cancer.

Stella V Paffenholz, Camilla Salvagno, Yu-Jui Ho, Matthew Limjoco, Timour Baslan, Sha Tian, Amanda Kulick, Elisa de Stanchina, John E Wilkinson, Francisco M Barriga, Dmitriy Zamarin, Juan R Cubillos-Ruiz, Josef Leibold, Scott W Lowe.

  High-grade serous ovarian carcinoma (HGSOC) is a cancer with dismal prognosis due to the limited effectiveness of existing chemo- and immunotherapies. To elucidate mechanisms mediating sensitivity or resistance to these therapies, we developed a fast and flexible autochthonous mouse model based on somatic introduction of HGSOC-associated genetic alterations into the ovary of immunocompetent mice using tissue electroporation. Tumors arising in these mice recapitulate the metastatic patterns and histological, molecular, and treatment response features of the human disease. By leveraging these models, we show that the ability to undergo senescence underlies the clinically observed increase in sensitivity of homologous recombination (HR)-deficient HGSOC tumors to platinum-based chemotherapy. Further, cGas/STING-mediated activation of a restricted senescence-associated secretory phenotype (SASP) was sufficient to induce immune infiltration and sensitize HR-deficient tumors to immune checkpoint blockade. In sum, our study identifies senescence propensity as a predictor of therapy response and defines a limited SASP profile that appears sufficient to confer added vulnerability to concurrent immunotherapy and, more broadly, provides a blueprint for the implementation of electroporation-based mouse models to reveal mechanisms of oncogenesis and therapy response in HGSOC.

Keywords:  cancer immunotherapy; mouse models; ovarian cancer; senescence

DOI:  https://doi.org/10.1073/pnas.2117754119
Cancer Res. 2022 Jan 25. pii: canres.CAN-21-2576-E.2021. [Epub ahead of print]

Hotspot ESR1 mutations are multimodal and contextual modulators of breast cancer metastasis.

Zheqi Li, Yang Wu, Megan E Yates, Nilgun Tasdemir, Amir Bahreini, Jian Chen, Kevin M Levine, Nolan M Priedigkeit, Azadeh Nasrazadani, Simak Ali, Laki Buluwela, Spencer Arnesen, Jason Gertz, Jennifer K Richer, Benjamin Troness, Dorraya El-Ashry, Qiang Zhang, Lorenzo Gerratana, Youbin Zhang, Massimo Cristofanilli, Maritza A Montanez, Prithu Sundd, Callen T Wallace, Simon C Watkins, Caterina Fumagalli, Elena Guerini-Rocco, Li Zhu, George C Tseng, Nikhil Wagle, Jason S Carroll, Paul Jank, Carsten Denkert, Maria M Karsten, Jens-Uwe Blohmer, Ben Ho Park, Peter C Lucas, Jennifer M Atkinson, Adrian V Lee, Steffi Oesterreich.

Constitutively active estrogen receptor-α (ER/ESR1) mutations have been identified in approximately one third of ER+ metastatic breast cancers. Although these mutations are known mediators of endocrine resistance, their potential role in promoting metastatic disease has not yet been mechanistically addressed. In this study, we show the presence of ESR1 mutations exclusively in distant but not local recurrences in five independent breast cancer cohorts. In concordance with transcriptomic profiling of ESR1 mutant tumors, genome-edited ESR1 Y537S and D538G mutant cell models exhibited a reprogrammed cell adhesive gene network via alterations in desmosome/gap junction genes and the TIMP3/MMP axis, which functionally conferred enhanced cell-cell contacts while decreasing cell-extracellular matrix (ECM) adhesion. In vivo studies showed ESR1 mutant cells were associated with larger multi-cellular circulating tumor cell (CTC) clusters with increased compactness compared to ESR1 WT CTCs. These preclinical findings translated to clinical observations, where CTC clusters were enriched in patients with ESR1-mutated metastatic breast cancer. Conversely, context-dependent migratory phenotypes revealed co-targeting of Wnt and ER as a vulnerability in a D538G cell model. Mechanistically, mutant ESR1 exhibited non-canonical regulation of several metastatic pathways, including secondary transcriptional regulation and de novo FOXA1-driven chromatin remodeling. Collectively, these data provide evidence for ESR1 mutation-modulated metastasis and suggest future therapeutic strategies for targeting ESR1 mutant breast cancer.

DOI: https://doi.org/10.1158/0008-5472.CAN-21-2576
Nat Commun. 2022 Jan 26. 13(1): 514

Mapping molecular subtype specific alterations in breast cancer brain metastases identifies clinically relevant vulnerabilities.

Nicola Cosgrove, Damir Varešlija, Stephen Keelan, Ashuvinee Elangovan, Jennifer M Atkinson, Sinéad Cocchiglia, Fiona T Bane, Vikrant Singh, Simon Furney, Chunling Hu, Jodi M Carter, Steven N Hart, Siddhartha Yadav, Matthew P Goetz, Arnold D K Hill, Steffi Oesterreich, Adrian V Lee, Fergus J Couch, Leonie S Young.

The molecular events and transcriptional plasticity driving brain metastasis in clinically relevant breast tumor subtypes has not been determined. Here we comprehensively dissect genomic, transcriptomic and clinical data in patient-matched longitudinal tumor samples, and unravel distinct transcriptional programs enriched in brain metastasis. We report on subtype specific hub genes and functional processes, central to disease-affected networks in brain metastasis. Importantly, in luminal brain metastases we identify homologous recombination deficiency operative in transcriptomic and genomic data with recurrent breast mutational signatures A, F and K, associated with mismatch repair defects, TP53 mutations and homologous recombination deficiency (HRD) respectively. Utilizing PARP inhibition in patient-derived brain metastatic tumor explants we functionally validate HRD as a key vulnerability. Here, we demonstrate a functionally relevant HRD evident at genomic and transcriptomic levels pointing to genomic instability in breast cancer brain metastasis which is of potential translational significance.

DOI: https://doi.org/10.1038/s41467-022-27987-5
Cancer Discov. 2022 Jan 25. pii: candisc.0910.2021. [Epub ahead of print]

Targeting S100A9-ALDH1A1-retinoic acid signaling to suppress brain relapse in EGFR-mutant lung cancer.

Anup Kumar Biswas, Seoyoung Han, Yifan Tai, Wanchao Ma, Courtney Coker, S Aidan Quinn, Ahmad Rushdi Shakri, Timothy James Zhong, Hanna Scholze, Galina G Lagos, Angeliki Mela, Katia Manova-Todorova, Elisa de Stanchina, Adolfo A Ferrando, Cathy Mendelsohn, Peter Canoll, Helena A Yu, Paul K Paik, Anjali Saqi, Catherine A Shu, Mark G Kris, Joan Massague, Swarnali Acharyya.

The epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) osimertinib has significantly prolonged progression-free survival (PFS) in EGFR-mutant lung cancer patients, including those with brain metastases. However, despite striking initial responses, osimertinib-treated patients eventually develop lethal metastatic relapse, often to the brain. Although osimertinib-refractory brain relapse is a major clinical challenge, its underlying mechanisms remain poorly understood. Using metastatic models of EGFR-mutant lung cancer, we show that cancer cells expressing high intracellular S100A9 escape osimertinib and initiate brain relapses. Mechanistically, S100A9 upregulates ALDH1A1 expression and activates the retinoic acid (RA) signaling pathway in osimertinib-refractory cancer cells. We demonstrate that the genetic repression of S100A9, ALDH1A1, or RA receptors (RAR) in cancer cells, or treatment with a pan-RAR antagonist, dramatically reduces brain metastasis. Importantly, S100A9 expression in cancer cells correlates with poor PFS in osimertinib-treated patients. Our study therefore identifies a novel, therapeutically targetable S100A9-ALDH1A1-RA axis that drives brain relapse.

DOI: https://doi.org/10.1158/2159-8290.CD-21-0910
J Clin Invest. 2022 Jan 27. pii: e152394. [Epub ahead of print]

Smad3 methylation by EZH2 promotes its activation and tumor metastasis.

Changsheng Huang, Fuqing Hu, Da Song, Xuling Sun, Anyi Liu, Qi Wu, Xiaowei She, Yaqi Chen, Lisheng Chen, Fayong Hu, Feng Xu, Xuelai Luo, Yongdong Feng, Xiangping Yang, Junbo Hu, Guihua Wang.

  SMAD3 plays a central role in cancer metastasis, and its hyperactivation is linked to poor cancer outcomes. Thus, it is critical to understand the upstream signaling pathways that govern SMAD3 activation. Here, we report that SMAD3 undergoes methylation at K53 and K333 by EZH2, a process crucial for cell membrane recruitment, phosphorylation, and activation of SMAD3 upon TGFB1 stimulation. Mechanistically, EZH2-triggered SMAD3 methylation facilitates SMAD3 interaction with its cellular membrane localization molecule (SARA), which in turn sustains SMAD3 phosphorylation by the TGFB receptor. Pathologically, EZH2 expression increasing results in the accumulation of SMAD3 methylation to facilitate SMAD3 activation. EZH2-mediated SMAD3 K53/K333 methylation is upregulated and correlated with SMAD3 hyperactivation in breast cancer, promotes tumor metastasis, and is predictive of poor survival outcome. We used two TAT-peptides to abrogate SMAD3 methylation and therapeutically inhibit cancer metastasis. Collectively, these findings reveal the complicated layers in regulation of SMAD3 activation coordinated by EZH2-mediated SMAD3 K53/K333 methylation to drive cancer metastasis.

Keywords:  Cancer; Molecular biology; Oncology

DOI:  https://doi.org/10.1172/JCI152394
Cell Death Differ. 2022 Jan 22.

IKBKE phosphorylates and stabilizes Snail to promote breast cancer invasion and metastasis.

Wei Xie, Qiwei Jiang, Xueji Wu, Lei Wang, Bing Gao, Zicheng Sun, Xiaomei Zhang, Lang Bu, Ying Lin, Qiang Huang, Jie Li, Jianping Guo.

IKBKE, a non-canonical inflammatory kinase, is frequently amplified or activated, and plays predominantly oncogenic roles in human cancers, especially in breast cancer. However, the potential function and underlying mechanism of IKBKE contributing to breast cancer metastasis remain largely elusive. Here, we report that depletion of Ikbke markedly decreases polyoma virus middle T antigen (PyVMT)-induced mouse mammary tumorigenesis and subsequent lung metastasis. Biologically, ectopic expression of IKBKE accelerates, whereas depletion of IKBKE attenuates breast cancer invasiveness and migration in vitro and tumor metastasis in vivo. Mechanistically, IKBKE tightly controls the stability of transcriptional factor Snail in different layers, in particular by directly phosphorylating Snail, which markedly blocks the E3 ligase β-TRCP1-mediated Snail degradation, resulting in breast cancer epithelial-mesenchymal transition (EMT) and metastasis. These findings together reveal a novel oncogenic function of IKBKE in promoting breast cancer metastasis by governing Snail abundance, and highlight the potential of targeting IKBKE for metastatic breast cancer therapies.

DOI: https://doi.org/10.1038/s41418-022-00940-1
Cancer Res. 2022 Jan 25. pii: canres.1327.2021. [Epub ahead of print]

Functional analysis of MET exon 14 skipping alteration in cancer invasion and metastatic dissemination.

Feng Wang, Yang Liu, Wanglong Qiu, Elaine Shum, Monica Feng, Dejian Zhao, Deyou Zheng, Alain Borczuk, Haiying Cheng, Balazs Halmos.

MET exon 14 skipping alteration (MET∆14Ex) is an actionable oncogenic driver that occurs in 2-4% of non-small cell lung cancer (NSCLC) cases. The precise role of MET∆14Ex in tumor progression of NSCLC is poorly understood. Using multiple isogenic MET∆14Ex cell models established with CRISPR editing, we demonstrate that MET∆14Ex expression increases receptor kinase activity and downstream signaling by impairing receptor internalization and endocytic degradation, significantly boosting cell scatter, migration, and invasion capacity in vitro as well as metastasis in vivo. RNA sequencing analysis revealed that MET∆14Ex preferentially activates biological processes associated with cell movement, providing novel insights into its unique molecular mechanism of action. Activation of PI3K/Akt/Rac1 signaling and upregulation of multiple matrix metallopeptidases (MMPs) by MET∆14Ex induced cytoskeleton remodeling and extracellular matrix disassembly, which are critical functional pathways that facilitate cell invasion and metastasis. Therapeutically, MET inhibitors dramatically repressed MET∆14Ex-mediated tumor growth and metastasis in vivo, indicating potential therapeutic options for MET∆14Ex-altered NSCLC patients. These mechanistic insights into MET∆14Ex-mediated invasion and metastasis provide a deeper understanding of the role of MET∆14Ex in NSCLC.

DOI: https://doi.org/10.1158/0008-5472.CAN-21-1327
Lancet Oncol. 2022 Jan 24. pii: S1470-2045(22)00022-5. [Epub ahead of print]

Medical treatment for active breast cancer brain metastases.

Caroline Bailleux, Thomas Bachelot.

DOI: https://doi.org/10.1016/S1470-2045(22)00022-5
Cancer Discov. 2022 Jan 27. pii: candisc.0932.2021. [Epub ahead of print]

Tumor ENPP1(CD203a)/Haptoglobin Axis Exploits Myeloid-Derived Suppressor Cells to Promote Post-Radiotherapy Local Recurrence in Breast Cancer.

Borja Ruiz-Fernandez de Cordoba, Haritz Moreno, Karmele Valencia, Naiara Perurena, Pablo Ruedas, Thomas Walle, Alberto Pezonaga-Torres, Juan Hinojosa, Elisabet Guruceaga, Antonio Pineda-Lucena, Marta Abengozar-Muela, Denis Cochonneau, Carolina Zandueta, Susana Martinez-Canarias, Alvaro Teijeira, Daniel Ajona, Sergio Ortiz-Espinosa, Xabier Morales, Carlos Ortiz de Solorzano, Marta Santisteban, Luis I Ramos-Garcia, Laura Guembe, Vratislav Strnad, Dominique Heymann, Sandra Hervas-Stubbs, Ruben Pio, Maria E Rodriguez-Ruiz, Carlos E de Andrea, Silvestre Vicent, Ignacio Melero, Fernando Lecanda, Rafael Martinez-Monge.

Locoregional failure (LRF) in breast cancer patients post-surgery and post-irradiation (IR) is linked to a dismal prognosis. In a refined new model, we identified Enpp1 (Ectonucleotide pyrophosphatase /phosphodiesterase 1/CD203a) to be closely associated with LRF. Enpp1high circulating tumor cells (CTC) contribute to relapse by a self-seeding mechanism. This process requires the infiltration of PMN-MDSC and neutrophil extracellular traps (NET) formation. Genetic and pharmacological Enpp1 inhibition or NET blockade extend relapse-free survival. Furthermore, in combination with fractionated irradiation (FD), Enpp1 abrogation obliterates LRF. Mechanistically, Enpp1-generated adenosinergic metabolites enhance Haptoglobin (Hp) expression. This inflammatory mediator elicits myeloid invasiveness and promotes NET formation. Accordingly, a significant increase in ENPP1 and NET formation is detected in relapsed human breast cancer tumors. Moreover, high ENPP1 or HP levels are associated with poor prognosis. These findings unveil the ENPP1/HP axis as an unanticipated mechanism exploited by tumor cells linking inflammation to immune remodeling favoring local relapse.

DOI: https://doi.org/10.1158/2159-8290.CD-21-0932
Cell Rep. 2022 Jan 25. pii: S2211-1247(22)00013-4. [Epub ahead of print]38(4): 110301

Endothelial p130cas confers resistance to anti-angiogenesis therapy.

Yunfei Wen, Anca Chelariu-Raicu, Sujanitha Umamaheswaran, Alpa M Nick, Elaine Stur, Pahul Hanjra, Dahai Jiang, Nicholas B Jennings, Xiuhui Chen, Sara Corvigno, Deanna Glassman, Gabriel Lopez-Berestein, Jinsong Liu, Mien-Chie Hung, Anil K Sood.

  Anti-angiogenic therapies, such as anti-VEGF antibodies (AVAs), have shown promise in clinical settings. However, adaptive resistance to such therapies occurs frequently. We use orthotopic ovarian cancer models with AVA-adaptive resistance to investigate the underlying mechanisms. Genomic profiling of AVA-resistant tumors guides us to endothelial p130cas. We find that bevacizumab induces cleavage of VEGFR2 in endothelial cells by caspase-10 and that VEGFR2 fragments internalize into the nucleus and autophagosomes. Nuclear VEGFR2 and p130cas fragments, together with TNKS1BP1 (tankyrase-1-binding protein), initiate endothelial cell death. Blockade of autophagy in AVA-resistant endothelial cells retains VEGFR2 at the membrane with bevacizumab treatment. Targeting host p130cas with RGD (Arg-Gly-Asp)-tagged nanoparticles or genomic ablation of vascular p130cas in p130casflox/floxTie2Cre mice significantly extends the survival of mice with AVA-resistant ovarian tumors. Higher vascular p130cas is associated with shorter survival of individuals with ovarian cancer. Our findings identify opportunities for new strategies to overcome adaptive resistance to AVA therapy.

Keywords:  TNKS1BP1; VEGFR2; adaptive resistance; angiogenesis; anti-angiogenic therapy; autophagy; p130cas

DOI:  https://doi.org/10.1016/j.celrep.2022.110301
Mol Cancer. 2022 Jan 24. 21(1): 29

The circROBO1/KLF5/FUS feedback loop regulates the liver metastasis of breast cancer by inhibiting the selective autophagy of afadin.

Zehao Wang, Lu Yang, Peng Wu, Xing Li, Yuhui Tang, Xueqi Ou, Yue Zhang, Xiangsheng Xiao, Jin Wang, Hailin Tang.

   BACKGROUND: Metastasis causes the majority of cancer-related deaths worldwide. Increasing studies have revealed that circRNAs are associated with the carcinogenesis and metastasis of many cancers. Nevertheless, the biological mechanisms of circRNAs in breast cancer (BC) liver metastasis remain extremely ambiguous.
METHODS: In this study, we identified circROBO1 from three pairs of primary BC and metastatic liver sites by RNA sequencing. FISH assays and RT-qPCR were conducted to validate the existence and expression of circROBO1. The oncogenic role of circROBO1 was demonstrated both in vitro and in vivo. Western blot, ChIP, RIP, RNA pull-down, and dual-luciferase reporter assays were used to confirm the interaction of the feedback loop among circROBO1, miR-217-5p, KLF5, and FUS. Meanwhile, the regulation of selective autophagy was investigated by immunofluorescence, CoIP, and western blot.
RESULTS: In this study, upregulated expression of circROBO1 was found in BC-derived liver metastases and was correlated with poor prognosis. Knockdown of circROBO1 strikingly inhibited the proliferation, migration, and invasion of BC cells, whereas overexpression of circROBO1 showed the opposite effects. Moreover, overexpression of circROBO1 promoted tumor growth and liver metastasis in vivo. Further research revealed that circROBO1 could upregulate KLF5 by sponging miR-217-5p, allowing KLF5 to activate the transcription of FUS, which would promote the back splicing of circROBO1. Therefore, a positive feedback loop comprising circROBO1/KLF5/FUS was formed. More importantly, we found that circROBO1 inhibited selective autophagy of afadin by upregulating KLF5.
CONCLUSIONS: Our results demonstrated that circROBO1 facilitates the carcinogenesis and liver metastasis of BC through the circROBO1/KLF5/FUS feedback loop, which inhibits the selective autophagy of afadin by suppressing the transcription of BECN1. Therefore, circROBO1 could be used not only as a potential prognostic marker but also as a therapeutic target in BC.

Keywords:  Afadin; Autophagy; BC; BECN1; FUS; KLF5; Metastasis; circROBO1; miR-217

DOI:  https://doi.org/10.1186/s12943-022-01498-9
Oncogene. 2022 Jan 27.

Targeting a splicing-mediated drug resistance mechanism in prostate cancer by inhibiting transcriptional regulation by PKCβ1.

James E Melnyk, Veronica Steri, Hao G Nguyen, Y Christina Hwang, John D Gordan, Byron Hann, Felix Y Feng, Kevan M Shokat.

The androgen receptor (AR) is a central driver of aggressive prostate cancer. After initial treatment with androgen receptor signaling inhibitors (ARSi), reactivation of AR signaling leads to resistance. Alternative splicing of AR mRNA yields the AR-V7 splice variant, which is currently an undruggable mechanism of ARSi resistance: AR-V7 lacks a ligand binding domain, where hormones and anti-androgen antagonists act, but still activates AR signaling. We reveal PKCβ as a druggable regulator of transcription and splicing at the AR genomic locus. We identify a clinical PKCβ inhibitor in combination with an FDA-approved anti-androgen as an approach for repressing AR genomic locus expression, including expression of AR-V7, while antagonizing full-length AR. PKCβ inhibition reduces total AR gene expression, thus reducing AR-V7 protein levels and sensitizing prostate cancer cells to current anti-androgen therapies. We demonstrate that this combination may be a viable therapeutic strategy for AR-V7-positive prostate cancer.

DOI: https://doi.org/10.1038/s41388-022-02179-z
Sci Transl Med. 2022 Jan 26. 14(629): eabh2548

Loss of miR-31-5p drives hematopoietic stem cell malignant transformation and restoration eliminates leukemia stem cells in mice.

Biying Zhu, Wenbin Zhong, Xiuye Cao, Guoping Pan, Mengyang Xu, Jie Zheng, Huanzhao Chen, Xiaoqin Feng, Chengwei Luo, Chen Lu, Jie Xiao, Weize Lin, Chaofeng Lai, Mingchuan Li, Xin Du, Qing Yi, Daoguang Yan.

Leukemia stem cells (LSCs) propagate leukemia and are responsible for the high frequency of relapse of treated patients. The ability to target LSCs remains elusive, indicating a need to understand the underlying mechanism of LSC formation. Here, we report that miR-31-5p is reduced or undetectable in human LSCs compared to hematopoietic stem progenitor cells (HSPCs). Inhibition of miR-31-5p in HSPCs promotes the expression of its target gene FIH, encoding FIH [factor inhibiting hypoxia-inducing factor 1α (HIF-1α)], to suppress HIF-1α signaling. Increased FIH resulted in a switch from glycolysis to oxidative phosphorylation (OXPHOS) as the predominant mode of energy metabolism and increased the abundance of the oncometabolite fumarate. Increased fumarate promoted the conversion of HSPCs to LSCs and initiated myeloid leukemia-like disease in NOD-Prkdcscid IL2rgtm1/Bcgen (B-NDG) mice. We further demonstrated that miR-31-5p inhibited long- and short-term hematopoietic stem cells with a high frequency of LSCs. In combination with the chemotherapeutic agent Ara-C (cytosine arabinoside), restoration of miR-31-5p using G7 poly (amidoamine) nanosized dendriplex encapsulating miR-31-5p eliminated LSCs and inhibited acute myeloid leukemia (AML) progression in patient-derived xenograft mouse models. These results demonstrated a mechanism of HSC malignant transformation through altered energy metabolism and provided a potential therapeutic strategy to treat patients with AML.

DOI: https://doi.org/10.1126/scitranslmed.abh2548
Gut. 2022 Jan 27. pii: gutjnl-2021-325137. [Epub ahead of print]

Tumour-associated neutrophils secrete AGR2 to promote colorectal cancer metastasis via its receptor CD98hc-xCT.

Shaobo Tian, Yanan Chu, Jia Hu, Xueliang Ding, Zhibo Liu, Daan Fu, Ye Yuan, Yan Deng, Guobin Wang, Lin Wang, Zheng Wang.

   OBJECTIVE: Reciprocal cellular crosstalk within the tumour microenvironment (TME) actively participates in tumour progression. The anterior gradient-2 (AGR2) can be secreted to extracellular compartments and contribute to colorectal cancer (CRC) metastasis. We investigated the cellular source for secreted AGR2 in the TME and underlying mechanisms mediating secreted AGR2's effects.
DESIGN: Tissue microarray, tumour tissues, blood samples and tumour-associated neutrophils (TANs) from patients with CRC were isolated for phenotypical and functional analyses. The role of TAN-secreted AGR2 was determined in neutrophil-specific Agr2 knockout (Agr2f/f;Mrp-Cre) mice. The biological roles and mechanisms of secreted AGR2 in CRC metastasis were determined in vitro and in vivo.
RESULTS: TANs were a predominant cell type for secreting AGR2 in the TME of CRC. TANs-secreted AGR2 promoted CRC cells' migration. Neutrophils-specific ablation of Agr2 in mice ameliorated CRC liver metastases. The heavy chain of CD98 (CD98hc) served as the functional receptor for secreted AGR2. Mechanistically, secreted AGR2 increased xCT activity in a CD98hc-dependent manner, subsequently activating Ras homologue family member A/Rho-associated protein kinase 2 cascade. CRC cells actively recruited TANs through the C-X-C motif chemokine 2. Moreover, CRC-derived transforming growth factor beta 1 (TGF-β1) educated peripheral blood neutrophils to become AGR2+ TANs that secrete AGR2. Abundant infiltration of AGR2+ TANs and high expression of TGF-β1 and CD98hc-xCT were correlated with poor prognosis of patients with CRC.
CONCLUSIONS: Our study unveils a novel crosstalk between TANs and CRC cells involving the secreted AGR2-CD98hc-xCT axis that promotes metastasis and impacts the outcomes of patients with CRC.

Keywords:  colorectal cancer; liver metastases

DOI:  https://doi.org/10.1136/gutjnl-2021-325137
Oncogene. 2022 Jan 24.

Canonical TGFβ signaling induces collective invasion in colorectal carcinogenesis through a Snail1- and Zeb1-independent partial EMT.

Marion Flum, Severin Dicks, Yu-Hsiang Teng, Monika Schrempp, Alexander Nyström, Melanie Boerries, Andreas Hecht.

Local invasion is the initial step towards metastasis, the main cause of cancer mortality. In human colorectal cancer (CRC), malignant cells predominantly invade as cohesive collectives and may undergo partial epithelial-mesenchymal transition (pEMT) at the invasive front. How this particular mode of stromal infiltration is generated is unknown. Here we investigated the impact of oncogenic transformation and the microenvironment on tumor cell invasion using genetically engineered organoids as CRC models. We found that inactivation of the Apc tumor suppressor combined with expression of oncogenic KrasG12D and dominant-negative Trp53R172H did not cell-autonomously induce invasion in vitro. However, oncogenic transformation primed organoids for activation of a collective invasion program upon exposure to the prototypical microenvironmental factor TGFβ1. Execution of this program co-depended on a permissive extracellular matrix which was further actively remodeled by invading organoids. Although organoids shed some epithelial properties particularly at the invasive edge, TGFβ1-stimulated organoids largely maintained epithelial gene expression while additionally implementing a mesenchymal transcription pattern, resulting in a pEMT phenotype that did not progress to a fully mesenchymal state. Notably, while TGFβ1 induced pEMT and promoted collective invasion, it abrogated self-renewal capacity of TKA organoids which correlated with the downregulation of intestinal stem cell (ISC) marker genes. Mechanistically, induction of the non-progressive pEMT required canonical TGFβ signaling mediated by Smad transcription factors (TFs), whereas the EMT master regulators Snail1 and Zeb1 were dispensable. Gene expression profiling provided further evidence for pEMT of TGFβ1-treated organoids and showed that their transcriptomes resemble those of human poor prognosis CMS4 cancers which likewise exhibit pEMT features. We propose that collective invasion in colorectal carcinogenesis is triggered by microenvironmental stimuli through activation of a novel, transcription-mediated form of non-progressive pEMT independently of classical EMT regulators.

DOI: https://doi.org/10.1038/s41388-022-02190-4
Cell Rep. 2022 Jan 25. pii: S2211-1247(21)01731-9. [Epub ahead of print]38(4): 110227

Disruption of pancreatic stellate cell myofibroblast phenotype promotes pancreatic tumor invasion.

Elizabeth R Murray, Shinelle Menezes, Jack C Henry, Josie L Williams, Lorena Alba-Castellón, Priththivika Baskaran, Ivan Quétier, Ami Desai, Jacqueline J T Marshall, Ian Rosewell, Marianthi Tatari, Vinothini Rajeeve, Faraz Khan, Jun Wang, Panoraia Kotantaki, Eleanor J Tyler, Namrata Singh, Claire S Reader, Edward P Carter, Kairbaan Hodivala-Dilke, Richard P Grose, Hemant M Kocher, Nuria Gavara, Oliver Pearce, Pedro Cutillas, John F Marshall, Angus J M Cameron.

  In pancreatic ductal adenocarcinoma (PDAC), differentiation of pancreatic stellate cells (PSCs) into myofibroblast-like cancer-associated fibroblasts (CAFs) can both promote and suppress tumor progression. Here, we show that the Rho effector protein kinase N2 (PKN2) is critical for PSC myofibroblast differentiation. Loss of PKN2 is associated with reduced PSC proliferation, contractility, and alpha-smooth muscle actin (α-SMA) stress fibers. In spheroid co-cultures with PDAC cells, loss of PKN2 prevents PSC invasion but, counter-intuitively, promotes invasive cancer cell outgrowth. PKN2 deletion induces a myofibroblast to inflammatory CAF switch in the PSC matrisome signature both in vitro and in vivo. Further, deletion of PKN2 in the pancreatic stroma induces more locally invasive, orthotopic pancreatic tumors. Finally, we demonstrate that a PKN2KO matrisome signature predicts poor outcome in pancreatic and other solid human cancers. Our data indicate that suppressing PSC myofibroblast function can limit important stromal tumor-suppressive mechanisms, while promoting a switch to a cancer-supporting CAF phenotype.

Keywords:  CAF; PKN2; Rho GTPases; cancer-associated fibroblasts; matrisome; pancreatic cancer; protein kinase N2; tumour microenvironment

DOI:  https://doi.org/10.1016/j.celrep.2021.110227
Ann Oncol. 2022 Jan 20. pii: S0923-7534(22)00009-6. [Epub ahead of print]

Single cell DNA-seq depicts clonal evolution of multiple driver alterations in osimertinib resistant patients.

J Chen, F Facchinetti, F Braye, A A Yurchenko, L Bigot, S Ponce, D Planchard, A Gazzah, S Nikolaev, S Michiels, D Vasseur, L Lacroix, L Tselikas, C Nobre, K A Olaussen, F Andre, J Y Scoazec, F Barlesi, J C Soria, Y Loriot, B Besse, L Friboulet.

   BACKGROUND: The development of targeted agents, such as osimertinib for EGFR-mutated NSCLC, has drastically improved patient outcome, but tumor resistance eventually always occurs. In osimertinib resistant NSCLC, the emergence of a second molecular driver alteration (such as ALK, RET, FGFR3 fusions or BRAF, KRAS mutations) has been described. Whether those alterations and the activating EGFR mutations occur within a single cancer cell or in distinct cell populations is largely debated.
PATIENTS AND METHODS: Tumor sequencing was used to identify the acquired resistance mechanisms to osimertinib in the MATCH-R trial (NCT0251782). We implemented single cell NGS to investigate tumor heterogeneity on patient's frozen tissues in which multiple alterations have been identified. Patient-derived models, cell lines and PDX, were exposed to specific inhibitors to investigate combination treatment strategies.
RESULTS: Among the 45 patients included in MATCH-R who progressed on osimertinib, nine developed a second targetable alteration (n = 2 FGFR3-TACC3, n = 1 KIF5B-RET, n = 1 STRN-ALK fusions; n = 2 BRAFV600E, n = 1 KRASG12V, n = 1 KRASG12R, n = 1 KRASG12D mutations). Single cell analysis revealed that the two driver alterations co-exist within one single cancer cell in the four patients whose frozen samples were fully contributive. A high degree of heterogeneity within samples and sequential acquisitions of molecular events were highlighted. A combination treatment concomitantly targeting the two driver alterations was required on the corresponding patient-derived models to restore cell sensitivity, which was consistent with clinical data showing efficacy of brigatinib in the patient with ALK fusion after progression to osimertinib and crizotinib administered sequentially.
CONCLUSIONS: Distinct molecular driver alterations at osimertinib resistance co-exist with initial EGFR mutations in single cancer cells. The clonal evolution of cancer cell populations emphasized their heterogeneity leading to osimertinib relapse. Combining two targeted treatments is effective to achieve clinical benefit.

Keywords:  Single cell; clonal evolution; double alterations; osimertinib; resistance

DOI:  https://doi.org/10.1016/j.annonc.2022.01.004
Cell Rep. 2022 Jan 25. pii: S2211-1247(21)01765-4. [Epub ahead of print]38(4): 110253

Opioid receptor signaling suppresses leukemia through both catalytic and non-catalytic functions of TET2.

Huanhuan Zhao, Jun Lu, Tong Yan, Fei Han, Jie Sun, Xiaolin Yin, Liting Chen, Chao Shen, Mark Wunderlich, Weina Yun, Lingling Yang, Liyun Chen, Dan Su, Stefan K Bohlander, Fudi Wang, James C Mulloy, Chong Li, Jianjun Chen, He Huang, Xi Jiang.

  Acute myeloid leukemia (AML) is a genetically heterogeneous and frequently fatal malignancy. The ten-eleven translocation (TET)-mediated DNA demethylation is known to be critically associated with AML pathogenesis. Through chemical compound screening, we find that the opioid receptor agonist, loperamide hydrochloride (OPA1), significantly suppresses AML cell viability. The potential therapeutic effects of opioid receptor agonists, especially OPA1, are verified in AML cells in vitro and mouse and human AML models in vivo. OPA1-induced activation of OPRM1 signaling enhances the transcription of TET2 and thus activates both catalytic-dependent and -independent functions of TET2. Notably, AMLs with TET2 mutations or chemotherapy resistance are sensitive to OPA1 as well. Our results reveal the OPRM1-TET2 regulatory axis in AML and suggest that opioid agonists, particularly OPA1, a US Food and Drug Administration (FDA)-approved antidiarrheal drug, have therapeutic potential in AML, especially in TET2-mutated and chemotherapy-resistant AMLs, which have a poor prognosis.

Keywords:  5hmC; AML; TET; acute myeloid leukemia; loperamide; opioid signaling; targeted therapy

DOI:  https://doi.org/10.1016/j.celrep.2021.110253
Eur Urol. 2022 Jan 19. pii: S0302-2838(21)02287-9. [Epub ahead of print]

Quality and Quantity: Evaluating Tumor Biology Alongside Novel Imaging on Diagnosis of Metastatic Hormone-sensitive Prostate Cancer.

Vanessa Di Lalla, Michael J Kucharczyk, Alexander W Wyatt, Felix Y Feng, Stephan Probst, Gwenaelle Gravis, Jonathan So, Fred Saad, Tamim Niazi.

Tumor biology may play an important role as an effective predictive biomarker that is complementary to functional imaging for metastatic hormone-sensitive prostate cancer.

DOI: https://doi.org/10.1016/j.eururo.2021.12.038
Clin Cancer Res. 2022 Jan 25. pii: clincanres.3764.2021. [Epub ahead of print]

FREQUENCY AND GENOMIC ASPECTS OF INTRINSIC RESISTANCE TO VISMODEGIB IN LOCALLY ADVANCED BASAL CELL CARCINOMA.

Andrey A Yurchenko, Oltin T Pop, Meriem Ighilahriz, Ismael Padioleau, Fatemeh Rajabi, Hayley J Sharpe, Nicolas Poulalhon, Brigitte Dreno, Amir Khammari, Marc Delord, Antonio Alberdi, Nadem Soufir, Maxime Battistella, Samia Mourah, Fanny Bouquet, Ariel Savina, Andrej Besse, Max Mendez-Lopez, Florent Grange, Sandrine Monestier, Laurent Mortier, Nicolas Meyer, Caroline Dutriaux, Caroline Robert, Philippe Saïag, Florian Herms, Jerome Lambert, Fred de Sauvage, Nicolas Dumaz, Lukas Flatz, Nicole Basset-Seguin, Sergey I Nikolaev.

PURPOSE: Vismodegib is approved for the treatment of locally advanced basal cell carcinoma (laBCC), but some cases demonstrate intrinsic resistance (IR) to the drug. We sought to assess the frequency of IR to vismodegib in laBCC and its underlying genomic mechanisms.
EXPERIMENTAL DESIGN: Response to vismodegib was evaluated in a cohort of 148 laBCC patients. Comprehensive genomic and transcriptomic profiling was performed in a subset of 5 intrinsically resistant BCC (IR-BCC).
RESULTS: We identified that IR-BCC represents 6.1% of laBCC in the studied cohort. Prior treatment with chemotherapy was associated with IR. Genetic events which were previously associated with acquired resistance (AR) in BCC or medulloblastoma were observed in 3 out of 5 IR-BCC. However, IR-BCCs were distinct by highly rearranged polyploid genomes. Functional analyses identified hyper-activation of the HIPPO-YAP and WNT pathways at RNA and protein levels in IR-BCC. In vitro assay on BCC cell line further confirmed that YAP1 overexpression increases cell proliferation rate.
CONCLUSIONS: IR to vismodegib is a rare event in laBCC. IR-BCCs frequently harbor resistance mutations in the Hh pathway, but also are characterized by hyper-activation of HIPPO-YAP and WNT pathways.

DOI: https://doi.org/10.1158/1078-0432.CCR-21-3764