bims-pimaco 2021-05-09 papers

on PI3K and MAPK signalling in colorectal cancer

Issue of 2021–05–09
23 papers selected by
Lucas B. Zeiger, CRUK Scotland Institute, Beatson Institute for Cancer Research

Targeting the KRAS α4-α5 allosteric interface inhibits pancreatic cancer tumorigenesis.
Wnt and Src signals converge on YAP-TEAD to drive intestinal regeneration.
PIK3C3 Inhibition Promotes Sensitivity to Colon Cancer Therapy by Inhibiting Cancer Stem Cells.
Therapeutically reprogrammed nutrient signalling enhances nanoparticulate albumin bound drug uptake and efficacy in KRAS-mutant cancer.
Combined Targeting of AKT and mTOR Synergistically Inhibits Formation of Primary Colorectal Carcinoma Tumouroids In Vitro: A 3D Tumour Model for Pre-therapeutic Drug Screening.
Structural analysis of the PTEN:P-Rex2 signaling complex reveals how cancer-associated mutations coordinate to hyperactivate Rac1.
Lipid metabolism in cancer: New perspectives and emerging mechanisms.
Clinical and prognostic features of patients with detailed RAS/BRAF-mutant colorectal cancer in Japan.
Development of BRAF V600E Mutation in NRAS Q61L Mutated Rectal Cancer.
Functional Specificity of the Members of the Sos Family of Ras-GEF Activators: Novel Role of Sos2 in Control of Epidermal Stem Cell Homeostasis.
Id4 Suppresses the Growth and Invasion of Colorectal Cancer HCT116 Cells through CK18-Related Inhibition of AKT and EMT Signaling.
Clinicopathological factors associated with tumour-specific mutation detection in plasma of patients with RAS- or BRAF-mutated metastatic colorectal cancer.
Targeting the PI3K and MAPK pathways to improve response to HER2-targeted therapies in HER2-positive gastric cancer.
Improved Treatment Outcomes by Using Patient Specific Drug Combinations in Mammalian Target of Rapamycin Activated Advanced Metastatic Cancers.
Gain and loss of function mutations in biological chemical reaction networks: a mathematical model with application to colorectal cancer cells.
RAG-ulating mTORC1 with amino acids.
A signalling cascade for Ral.
Taurine Attenuates the Hypotaurine-Induced Progression of CRC via ERK/RSK Signaling.
Discordance of KRAS Mutational Status between Primary Tumors and Liver Metastases in Colorectal Cancer: Impact on Long-Term Survival Following Radical Resection.
Regulation of PCTAIRE1 protein stability by AKT1, LKB1, and BRCA1.
Breaking Oncogene Addiction: Getting RTK/RAS-Mutated Cancers off the SOS.
The genomic landscape of carcinomas with mucinous differentiation.
mTOR signaling regulates gastric epithelial progenitor homeostasis and gastric tumorigenesis via MEK1-ERKs and BMP-Smad1 pathways.

Small GTPases. 2021 May 05. 1-14

Targeting the KRAS α4-α5 allosteric interface inhibits pancreatic cancer tumorigenesis.

Imran Khan, Catherine MarElia-Bennet, Julia Lefler, Mariyam Zuberi, Eric Denbaum, Akiko Koide, Dean M Connor, Ann-Marie Broome, Thierry Pécot, Cynthia Timmers, Michael C Ostrowski, Shohei Koide, John P O'Bryan.

  RAS is the most frequently mutated oncogene in human cancer with nearly ~20% of cancer patients possessing mutations in one of three RAS genes (K, N or HRAS). However, KRAS is mutated in nearly 90% of pancreatic ductal carcinomas (PDAC). Although pharmacological inhibition of RAS has been challenging, KRAS(G12C)-specific inhibitors have recently entered the clinic. While KRAS(G12C) is frequently expressed in lung cancers, it is rare in PDAC. Thus, more broadly efficacious RAS inhibitors are needed for treating KRAS mutant-driven cancers such as PDAC. A RAS-specific tool biologic, NS1 Monobody, inhibits HRAS- and KRAS-mediated signalling and oncogenic transformation both in vitro and in vivo by targeting the α4-α5 allosteric site of RAS and blocking RAS self-association. Here, we evaluated the efficacy of targeting the α4-α5 interface of KRAS as an approach to inhibit PDAC development using an immunocompetent orthotopic mouse model. Chemically regulated NS1 expression inhibited ERK and AKT activation in KRAS(G12D) mutant KPC PDAC cells and reduced the formation and progression of pancreatic tumours. NS1-expressing tumours were characterized by increased infiltration of CD4 + T helper cells. These results suggest that targeting the #x3B1;4-#x3B1;5 allosteric site of KRAS may represent a viable therapeutic approach for inhibiting KRAS-mutant pancreatic tumours.

Keywords:  Monobody; Pancreatic ductal adenocarcinoma; T-cell; tumourigenesis

DOI:  https://doi.org/10.1080/21541248.2021.1906621
EMBO J. 2021 May 05. e105770

Wnt and Src signals converge on YAP-TEAD to drive intestinal regeneration.

Oriane Guillermin, Nikolaos Angelis, Clara M Sidor, Rachel Ridgway, Anna Baulies, Anna Kucharska, Pedro Antas, Melissa R Rose, Julia Cordero, Owen Sansom, Vivian S W Li, Barry J Thompson.

  Wnt signalling induces a gradient of stem/progenitor cell proliferation along the crypt-villus axis of the intestine, which becomes expanded during intestinal regeneration or tumour formation. The YAP transcriptional co-activator is known to be required for intestinal regeneration, but its mode of regulation remains controversial. Here we show that the YAP-TEAD transcription factor is a key downstream effector of Wnt signalling in the intestine. Loss of YAP activity by Yap/Taz conditional knockout results in sensitivity of crypt stem cells to apoptosis and reduced cell proliferation during regeneration. Gain of YAP activity by Lats1/2 conditional knockout is sufficient to drive a crypt hyperproliferation response. In particular, Wnt signalling acts transcriptionally to induce YAP and TEAD1/2/4 expression. YAP normally localises to the nucleus only in crypt base stem cells, but becomes nuclear in most intestinal epithelial cells during intestinal regeneration after irradiation, or during organoid growth, in a Src family kinase-dependent manner. YAP-driven crypt expansion during regeneration involves an elongation and flattening of the Wnt signalling gradient. Thus, Wnt and Src-YAP signals cooperate to drive intestinal regeneration.

Keywords:  Src; Wnt; YAP; intestine; regeneration

DOI:  https://doi.org/10.15252/embj.2020105770
Cancers (Basel). 2021 Apr 30. pii: 2168. [Epub ahead of print]13(9):

PIK3C3 Inhibition Promotes Sensitivity to Colon Cancer Therapy by Inhibiting Cancer Stem Cells.

Balawant Kumar, Rizwan Ahmad, Swagat Sharma, Saiprasad Gowrikumar, Mark Primeaux, Sandeep Rana, Amarnath Natarajan, David Oupicky, Corey R Hopkins, Punita Dhawan, Amar B Singh.

  Background: Despite recent advances in therapies, resistance to chemotherapy remains a critical problem in the clinical management of colorectal cancer (CRC). Cancer stem cells (CSCs) play a central role in therapy resistance. Thus, elimination of CSCs is crucial for effective CRC therapy; however, such strategies are limited. Autophagy promotes resistance to cancer therapy; however, whether autophagy protects CSCs to promote resistance to CRC-therapy is not well understood. Moreover, specific and potent autophagy inhibitors are warranted as clinical trials with hydroxychloroquine have not been successful. Methods: Colon cancer cells and tumoroids were used. Fluorescent reporter-based analysis of autophagy flux, spheroid and side population (SP) culture, and qPCR were done. We synthesized 36-077, a potent inhibitor of PIK3C3/VPS34 kinase, to inhibit autophagy. Combination treatments were done using 5-fluorouracil (5-FU) and 36-077. Results: The 5-FU treatment induced autophagy only in a subset of the treated colon cancer. These autophagy-enriched cells also showed increased expression of CSC markers. Co-treatment with 36-077 significantly improved efficacy of the 5-FU treatment. Mechanistic studies revealed that combination therapy inhibited GSK-3β/Wnt/β-catenin signaling to inhibit CSC population. Conclusion: Autophagy promotes resistance to CRC-therapy by specifically promoting GSK-3β/Wnt/β-catenin signaling to promote CSC survival, and 36-077, a PIK3C3/VPS34 inhibitor, helps promote efficacy of CRC therapy.

Keywords:  5-FloroUracil; PI3KC3; autophagy; cancer stem cells; chemoresistance

DOI:  https://doi.org/10.3390/cancers13092168
Nat Nanotechnol. 2021 May 06.

Therapeutically reprogrammed nutrient signalling enhances nanoparticulate albumin bound drug uptake and efficacy in KRAS-mutant cancer.

Ran Li, Thomas S C Ng, Stephanie J Wang, Mark Prytyskach, Christopher B Rodell, Hannes Mikula, Rainer H Kohler, Michelle A Garlin, Douglas A Lauffenburger, Sareh Parangi, Daniela M Dinulescu, Nabeel Bardeesy, Ralph Weissleder, Miles A Miller.

Nanoparticulate albumin bound paclitaxel (nab-paclitaxel, nab-PTX) is among the most widely prescribed nanomedicines in clinical use, yet it remains unclear how nanoformulation affects nab-PTX behaviour in the tumour microenvironment. Here, we quantified the biodistribution of the albumin carrier and its chemotherapeutic payload in optically cleared tumours of genetically engineered mouse models, and compared the behaviour of nab-PTX with other clinically relevant nanoparticles. We found that nab-PTX uptake is profoundly and distinctly affected by cancer-cell autonomous RAS signalling, and RAS/RAF/MEK/ERK inhibition blocked its selective delivery and efficacy. In contrast, a targeted screen revealed that IGF1R kinase inhibitors enhance uptake and efficacy of nab-PTX by mimicking glucose deprivation and promoting macropinocytosis via AMPK, a nutrient sensor in cells. This study thus shows how nanoparticulate albumin bound drug efficacy can be therapeutically improved by reprogramming nutrient signalling and enhancing macropinocytosis in cancer cells.

DOI: https://doi.org/10.1038/s41565-021-00897-1
Anticancer Res. 2021 May;41(5): 2257-2275

Combined Targeting of AKT and mTOR Synergistically Inhibits Formation of Primary Colorectal Carcinoma Tumouroids In Vitro: A 3D Tumour Model for Pre-therapeutic Drug Screening.

Dominik Nörz, Christina S Mullins, Daniel J Smit, Michael Linnebacher, Grith Hagel, Alp Mirdogan, Jacqueline Siekiera, Patrick Ehm, Jakob R Izbicki, Andreas Block, Ole Thastrup, Manfred Jücker.

   BACKGROUND: Pre-therapeutic analysis of three-dimensional spheroid cultures of primary tumour samples is a promising approach of assessing susceptibility to potential treatment. The phosphatidylinositol-3-kinase/AKT serine/threonine kinase/mammalian target of rapamycin (PI3K/AKT/mTOR) signalling pathway is frequently activated in colorectal cancer (CRC). In previous work, we showed combined inhibition of AKT and mTOR to be highly synergistic in cell lines from patients with hepatocellular carcinoma and cholangiocarcinoma in vitro as well as in vivo in murine xenograft tumour models.
MATERIALS AND METHODS: Patient-derived xenograft colorectal carcinoma cell lines HROC80 T1 M1, HROC147 T0 M1, HROC147Met, HROC277 T0 M1 and HROC277Met2 were treated with AKT inhibitor MK2206, mTOR inhibitor RAD001 or the combination of both drugs. The sensitivity of these cell lines to inhibition was evaluated by calculation of combinatory indices after bromodeoxyuridine assays and analysis of the respective pathways by western blotting. Furthermore, the dual inhibition of AKT and mTOR was confirmed in vivo in a xenograft mouse model. Additionally, primary CRC samples of four patients were embedded in a three-dimensional matrix and the sensitivity of these samples was analyzed by measurement of the spheroid area.
RESULTS: In this study, we demonstrate that combined treatment with MK2206 and RAD001 resulted in strong synergistic effects on growth of several primary CRC cell lines and reduced the growth of a patient-derived CRC xenograft in a xenotransplantation mouse model in vivo. Interestingly, the response to treatment varied between cell lines derived from the primary lesion and a liver metastasis of the same patient. In addition, combined treatment with AKT and mTOR inhibitors resulted in a synergistic inhibition of tumouroid growth in all four of the primary patient samples, analyzed in a three-dimensional spheroid model in vitro.
CONCLUSION: Our data demonstrate that combined treatment with AKT and mTOR inhibitors exhibits synergistic effects on proliferation of cell lines and primary tumour cells from patients with CRC and may be a promising approach for the treatment of CRC.

Keywords:  AKT; Colorectal cancer; combined treatment; mTOR; personalized medicine; pre-therapeutic drug screening; tumour spheroids; tumouroids

DOI:  https://doi.org/10.21873/anticanres.15002
Sci Signal. 2021 May 04. pii: eabc4078. [Epub ahead of print]14(681):

Structural analysis of the PTEN:P-Rex2 signaling complex reveals how cancer-associated mutations coordinate to hyperactivate Rac1.

Laura D'Andrea, Christina M Lucato, Elsa A Marquez, Yong-Gang Chang, Srgjan Civciristov, Chantel Mastos, Christopher J Lupton, Cheng Huang, Hans Elmlund, Ralf B Schittenhelm, Christina A Mitchell, James C Whisstock, Michelle L Halls, Andrew M Ellisdon.

The dual-specificity phosphatase PTEN functions as a tumor suppressor by hydrolyzing PI(3,4,5)P3 to PI(4,5)P2 to inhibit PI3K-AKT signaling and cellular proliferation. P-Rex2 is a guanine nucleotide exchange factor for Rho GTPases and can be activated by Gβγ subunits downstream of G protein-coupled receptor signaling and by PI(3,4,5)P3 downstream of receptor tyrosine kinases. The PTEN:P-Rex2 complex is a commonly mutated signaling node in metastatic cancer. Assembly of the PTEN:P-Rex2 complex inhibits the activity of both proteins, and its dysregulation can drive PI3K-AKT signaling and cellular proliferation. Here, using cross-linking mass spectrometry and functional studies, we gained mechanistic insights into PTEN:P-Rex2 complex assembly and coinhibition. We found that PTEN was anchored to P-Rex2 by interactions between the PDZ-interacting motif in the PTEN C-terminal tail and the second PDZ domain of P-Rex2. This interaction bridged PTEN across the P-Rex2 surface, preventing PI(3,4,5)P3 hydrolysis. Conversely, PTEN both allosterically promoted an autoinhibited conformation of P-Rex2 and blocked its binding to Gβγ. In addition, we observed that the PTEN-deactivating mutations and P-Rex2 truncations combined to drive Rac1 activation to a greater extent than did either single variant alone. These insights enabled us to propose a class of gain-of-function, cancer-associated mutations within the PTEN:P-Rex2 interface that uncouple PTEN from the inhibition of Rac1 signaling.

DOI: https://doi.org/10.1126/scisignal.abc4078
Dev Cell. 2021 May 03. pii: S1534-5807(21)00322-1. [Epub ahead of print]

Lipid metabolism in cancer: New perspectives and emerging mechanisms.

Lindsay A Broadfield, Antonino Alejandro Pane, Ali Talebi, Johannes V Swinnen, Sarah-Maria Fendt.

  Tumors undergo metabolic transformations to sustain uncontrolled proliferation, avoid cell death, and seed in secondary organs. An increased focus on cancer lipid metabolism has unveiled a number of mechanisms that promote tumor growth and survival, many of which are independent of classical cellular bioenergetics. These mechanisms include modulation of ferroptotic-mediated cell death, support during tumor metastasis, and interactions with the cells of the tumor microenvironment. As such, targeting lipid metabolism for anti-cancer therapies is attractive, with recent work on small-molecule inhibitors identifying compounds to target lipid metabolism. Here, we discuss these topics and identify open questions.

Keywords:  cancer; immunometabolism; lipids; metabolism; metastasis; tumor microenvironment

DOI:  https://doi.org/10.1016/j.devcel.2021.04.013
BMC Cancer. 2021 May 07. 21(1): 518

Clinical and prognostic features of patients with detailed RAS/BRAF-mutant colorectal cancer in Japan.

Tatsuki Ikoma, Mototsugu Shimokawa, Masahito Kotaka, Toshihiko Matsumoto, Hiroki Nagai, Shogen Boku, Nobuhiro Shibata, Hisateru Yasui, Hironaga Satake.

   BACKGROUND: RAS/BRAFV600E mutations are the most remarkable oncogenic driver mutations in colorectal cancer (CRC) and play an important role in treatment selection. No data are available regarding the clinical and prognostic features of patients with detailed RAS/BRAFV600E-mutant metastatic CRC (mCRC) in Japan.
METHODS: A total of 152 chemotherapy-naïve patients with mCRC were included in this study between August 2018 and July 2019. Tumor samples were collected, and RAS/BRAFV600E status was investigated. RAS/BRAFV600E status was examined using a MEBGEN RASKET-B kit and polymerase chain reaction reverse sequence-specific oligonucleotide method.
RESULTS: RAS/BRAFV600E mutations were detected in 54% of cases (KRAS codon 12, 26%; KRAS codon 13, 17%; KRAS non-Exon2, 5%; NRAS, 5%; and BRAFV600E, 7%). BRAFV600E-mutant CRC mainly existed in the right colon, whereas KRAS non-Exon2 and NRAS-mutant CRC was predominantly present in the left colon. KRAS non-Exon2 and NRAS-mutant CRC were associated with shorter survival time than RAS wild-type CRC (hazard ratio [HR], 2.26; 95% confidence interval [CI], 0.64-8.03; p = 0.19; HR, 2.42; 95% CI, 0.68-8.61; p = 0.16) and significantly shorter overall survival than KRAS Exon2-mutant CRC (HR, 3.88; 95% CI, 0.92-16.3; p = 0.04; HR, 4.80; 95% CI, 1.14-20.2; p = 0.02).
CONCLUSIONS: In our multicenter study, the findings elucidated the clinical and prognostic features of patients with detailed RAS/BRAFV600E-mutant mCRC in Japan.

Keywords:  Colorectal cancer; KRAS Exon2; KRAS non-Exon2; NRAS

DOI:  https://doi.org/10.1186/s12885-021-08271-z
Ann Clin Lab Sci. 2021 Mar;51(2): 262-266

Development of BRAF V600E Mutation in NRAS Q61L Mutated Rectal Cancer.

Mei-Na Cao, Kai-Ting Chen, Quan-Fu Li, Li Wang, Gao-Wa Jin, Ren-Tuya Na, Jing Ma, Yu-Ting Liu, Jin-Liang Gao.

BRAF and NRAS are oncogenes in the RAS/RAF/MEK/MAP-kinase signaling pathway. Coexistent mutations of BRAF and NRAS in a single colorectal cancer patient have always been considered mutually exclusive or at least rare. The clinical outcome of these patients remains undetermined. Herein we report a 53-year-old man harboring an NRAS Q61L mutation in his primary rectal carcinoma, who presented with a concomitant mutation of BRAF V600E in his liver metastasis biopsy 55 months after the primary CRC surgical resection. Our findings suggest that a BRAF and NRAS developed co-mutation may lead to a distinct clinicopathological progression. BRAF-mutated CRCwill not benefit from anti-RAS targeted therapy.

Keywords: BRAF; NRAS; colorectal cancer; concomitant mutation
Cancers (Basel). 2021 Apr 29. pii: 2152. [Epub ahead of print]13(9):

Functional Specificity of the Members of the Sos Family of Ras-GEF Activators: Novel Role of Sos2 in Control of Epidermal Stem Cell Homeostasis.

Fernando C Baltanás, Cynthia Mucientes-Valdivieso, L Francisco Lorenzo-Martín, Natalia Fernández-Parejo, Rósula García-Navas, Carmen Segrelles, Nuria Calzada, Rocío Fuentes-Mateos, Jesús M Paramio, Xosé R Bustelo, Eugenio Santos.

  Prior reports showed the critical requirement of Sos1 for epithelial carcinogenesis, but the specific functionalities of the homologous Sos1 and Sos2 GEFs in skin homeostasis and tumorigenesis remain unclear. Here, we characterize specific mechanistic roles played by Sos1 or Sos2 in primary mouse keratinocytes (a prevalent skin cell lineage) under different experimental conditions. Functional analyses of actively growing primary keratinocytes of relevant genotypes-WT, Sos1-KO, Sos2-KO, and Sos1/2-DKO-revealed a prevalent role of Sos1 regarding transcriptional regulation and control of RAS activation and mechanistic overlapping of Sos1 and Sos2 regarding cell proliferation and survival, with dominant contribution of Sos1 to the RAS-ERK axis and Sos2 to the RAS-PI3K/AKT axis. Sos1/2-DKO keratinocytes could not grow under 3D culture conditions, but single Sos1-KO and Sos2-KO keratinocytes were able to form pseudoepidermis structures that showed disorganized layer structure, reduced proliferation, and increased apoptosis in comparison with WT 3D cultures. Remarkably, analysis of the skin of both newborn and adult Sos2-KO mice uncovered a significant reduction of the population of stem cells located in hair follicles. These data confirm that Sos1 and Sos2 play specific, cell-autonomous functions in primary keratinocytes and reveal a novel, essential role of Sos2 in control of epidermal stem cell homeostasis.

Keywords:  GEF; KO; Ras signaling; Sos1; Sos2; keratinocytes; skin homeostasis

DOI:  https://doi.org/10.3390/cancers13092152
J Oncol. 2021 ;2021 6660486

Id4 Suppresses the Growth and Invasion of Colorectal Cancer HCT116 Cells through CK18-Related Inhibition of AKT and EMT Signaling.

Hui-Jing Chen, Yue Yu, Yan-Xia Sun, Chuan-Zhong Huang, Jie-Yu Li, Fang Liu, Guo-Xiang Guo, Yun-Bin Ye.

Id4 is one of the inhibitors of DNA-binding proteins (Id) and involved in the pathogenesis of numerous cancers. The specific mechanism underlying the Id4-mediated regulation of proliferation, invasion, and metastasis of colorectal cancer (CRC) cells is still largely unclear. In the present study, results showed CRC cells had a lower baseline Id4 expression than normal intestinal epithelial NCM460 cells. In order to explore the role of Id4 in the tumorigenicity, CRC HCT116 cells with stable Id4 expression were used, and results showed Id4 overexpression arrested the cell cycle at the G0/G1 phase, inhibited the cell proliferation and the colony formation, as well as suppressed the migration and invasion. In the in vivo model, Id4 overexpression inhibited the tumor growth and metastasis in the nude mice. Furthermore, Id4 overexpression upregulated the expression of proteins associated with cell proliferation, inhibited the PI3K/AKT pathway, and suppressed epithelial-mesenchymal transition (EMT) of HCT116 cells. Moreover, Id4 significantly decreased cytokeratin 18 (CK18) expression, but CK18 overexpression in Id4 expressing HCT116-Id4 cells rescued the activation of AKT, p-AKT, MMP2, MMP7, and E-cadherin. Collectively, our study indicated Id4 may inhibit CRC growth and metastasis through inhibiting the PI3K/AKT pathway in a CK18-dependent manner and suppressing EMT. Id4 may become a target for the treatment of CRC.

DOI: https://doi.org/10.1155/2021/6660486
Int J Cancer. 2021 May 07.

Clinicopathological factors associated with tumour-specific mutation detection in plasma of patients with RAS- or BRAF-mutated metastatic colorectal cancer.

Julian Hamfjord, Tormod Kyrre Guren, Bengt Glimelius, Halfdan Sorbye, Per Pfeiffer, Olav Dajani, Ole Christian Lingjaerde, Kjell Magne Tveit, Niels Pallisgaard, Karen-Lise Garm Spindler, Elin H Kure.

  Detection of tumour-specific circulating cell-free DNA in plasma (ctDNA) fails in a significant number of cases depending on the clinical context. The primary aim was to investigate clinicopathological factors associated with detection of ctDNA in patients with RAS-/BRAF-mutated metastatic colorectal cancer (mCRC) prior to first-line therapy. A secondary aim was to evaluate the prognostic impact of ctDNA compared to other biomarkers. Patients were included from the NORDIC-VII study (N = 253). ctDNA was sampled prior to treatment and analysed for hotspot tissue mutations (KRAS, NRAS and BRAF) using droplet digital PCR. Multivariable regression models were constructed to predict the probability of mutation detection and survival. Increasing radiological size of target lesions by increments of 1 cm (odds ratio (OR) = 1.18; 95% confidence interval (CI) 1.09-1.27; P < 0.001), intact primary tumour (OR = 3.17; 95% CI 1.22-8.22; P = 0.018) and more than one metastatic site (OR = 3.08; 95% CI 1.32-7.19; P = 0.009) were associated with mutation detection in plasma. Metastatic involvement of the lung was associated with non-detection (OR = 0.26; 95% CI 0.12-0.58; P = 0.001). Pre-analytical and analytical factors modulated detection. High allele frequencies of ctDNA indicated poor prognosis independently of CEA and CA19-9 (hazard ratio (HR) = 2.38; 95% CI 1.74-3.26; P < 0.001; N = 206). Clinicopathological characteristics should be carefully considered when evaluating ctDNA results from mCRC patients, especially when confronted with a plasma negative result. ctDNA may prove to be a clinically useful marker in the evaluation of mCRC treatment.

Keywords:  Colorectal cancer; blood biomarkers; carbohydrate antigen 19-9; carcinoembryonic antigen; tumour-specific circulating cell-free DNA

DOI:  https://doi.org/10.1002/ijc.33672
J Transl Med. 2021 May 01. 19(1): 184

Targeting the PI3K and MAPK pathways to improve response to HER2-targeted therapies in HER2-positive gastric cancer.

M Janusz Mezynski, Angela M Farrelly, Mattia Cremona, Aoife Carr, Clare Morgan, Julie Workman, Paul Armstrong, Jennifer McAuley, Stephen Madden, Joanna Fay, Katherine M Sheehan, Elaine W Kay, Ciara Holohan, Yasir Elamin, Shereen Rafee, Patrick G Morris, Oscar Breathnach, Liam Grogan, Bryan T Hennessy, Sinead Toomey.

   BACKGROUND: Aberrant PI3K signalling is implicated in trastuzumab resistance in HER2-positive gastric cancer (GC). The role of PI3K or MEK inhibitors in sensitising HER2-positive GCs to trastuzumab or in overcoming trastuzumab resistance is unclear.
METHODS: Using mass spectrometry-based genotyping we analysed 105 hotspot, non-synonymous somatic mutations in PIK3CA and ERBB-family (EGFR, ERBB2, ERBB3 and ERBB4) genes in gastric tumour samples from 69 patients. A panel of gastric cell lines (N87, OE19, ESO26, SNU16, KATOIII) were profiled for anti-proliferative response to the PI3K inhibitor copanlisib and the MEK1/2 inhibitor refametinib alone and in combination with anti-HER2 therapies.
RESULTS: Patients with HER2-positive GC had significantly poorer overall survival compared to HER2-negative patients (15.9 months vs. 35.7 months). Mutations in PIK3CA were only identified in HER2-negative tumours, while ERBB-family mutations were identified in HER2-positive and HER2-negative tumours. Copanlisib had anti-proliferative effects in 4/5 cell lines, with IC50s ranging from 23.4 (N87) to 93.8 nM (SNU16). All HER2-positive cell lines except SNU16 were sensitive to lapatinib (IC50s 0.04 µM-1.5 µM). OE19 cells were resistant to trastuzumab. The combination of lapatinib and copanlisib was synergistic in ESO-26 and OE-19 cells (ED50: 0.83 ± 0.19 and 0.88 ± 0.13, respectively) and additive in NCI-N87 cells (ED50:1.01 ± 0.55). The combination of copanlisib and trastuzumab significantly improved growth inhibition compared to either therapy alone in NCI-N87, ESO26 and OE19 cells (p < 0.05).
CONCLUSIONS: PI3K or MEK inhibition alone or in combination with anti-HER2 therapy may represent an improved treatment strategy for some patients with HER2-positive GC, and warrants further investigation in a clinical trial setting.

Keywords:  HER2-positive gastric cancer; MAPK; PI3K; Signalling pathway activation; Somatic mutations; Targeted therapies; Treatment resistance

DOI:  https://doi.org/10.1186/s12967-021-02842-1
Front Pharmacol. 2021 ;12 631135

Improved Treatment Outcomes by Using Patient Specific Drug Combinations in Mammalian Target of Rapamycin Activated Advanced Metastatic Cancers.

Timothy Crook, Darshana Patil, Andrew Gaya, Nicholas Plowman, Sewanti Limaye, Anantbhushan Ranade, Amit Bhatt, Raymond Page, Dadasaheb Akolkar.

  Background: Activation of the mTOR signaling pathway is ubiquitous in cancers and a favourable therapeutic target. However, presently approved mTOR inhibitor monotherapies have modest benefits in labeled indications while poor outcomes have been reported for mTOR inhibitor monotherapy when administered in a label-agnostic setting based on univariate molecular indications. The present study aimed to determine whether patient-specific combination regimens with mTOR inhibitors and other anticancer agents selected based on multi-analyte molecular and functional tumor interrogation (ETA: Encyclopedic Tumor Analysis) yields significant treatment response and survival benefits in advanced or refractory solid organ cancers. Methods: We evaluated treatment outcomes in 49 patients diagnosed with unresectable or metastatic solid organ cancers, of whom 3 were therapy naïve and 46 were pre-treated in whom the cancer had progressed on 2 or more prior systemic lines. All patients received mTOR inhibitor in combination with other targeted, endocrine or cytotoxic agents as guided by ETA. Patients were followed-up to determine Objective Response Rate (ORR), Progression Free Survival (PFS) and Overall Survival (OS). Results: The Objective Response Rate (ORR) was 57.1%, the disease Control rate (DCR) was 91.8%, median Progression Free Survival (mPFS) was 4.9 months and median Overall Survival (mOS) was 9.4 months. There were no Grade IV treatment related adverse events (AEs) or any treatment related deaths. Conclusion: Patient-specific combination regimens with mTOR inhibition and other anti-neoplastic agents, when selected based on multi-analyte molecular and functional profiling of the tumor can yield meaningful outcomes in advanced or refractory solid organ cancers. Trial Registration: Details of all trials are available at WHO-ICTRP: https://apps.who.int/trialsearch/. RESILIENT ID CTRI/2018/02/011808. ACTPRO ID CTRI/2018/05/014178. LIQUID IMPACT ID CTRI/2019/02/017548.

Keywords:  Akt; ETA; PI3K; PIK3CA; encyclopedic tumor analysis; mTOR; mTOR inhibitor; rapalog

DOI:  https://doi.org/10.3389/fphar.2021.631135
J Math Biol. 2021 May 04. 82(6): 55

Gain and loss of function mutations in biological chemical reaction networks: a mathematical model with application to colorectal cancer cells.

Sara Sommariva, Giacomo Caviglia, Michele Piana.

  This paper studies a system of Ordinary Differential Equations modeling a chemical reaction network and derives from it a simulation tool mimicking Loss of Function and Gain of Function mutations found in cancer cells. More specifically, from a theoretical perspective, our approach focuses on the determination of moiety conservation laws for the system and their relation with the corresponding stoichiometric surfaces. Then we show that Loss of Function mutations can be implemented in the model via modification of the initial conditions in the system, while Gain of Function mutations can be implemented by eliminating specific reactions. Finally, the model is utilized to examine in detail the G1-S phase of a colorectal cancer cell.

Keywords:  Colorectal cancer cells; G1-S transition point; Gain of function mutations; Loss of function mutations; Reaction kinetics; Synthetic cell biology

DOI:  https://doi.org/10.1007/s00285-021-01607-0
Nat Rev Mol Cell Biol. 2021 May 04.

RAG-ulating mTORC1 with amino acids.

Jenna L Jewell.

DOI: https://doi.org/10.1038/s41580-021-00378-2
Small GTPases. 2021 May 06. 1-8

A signalling cascade for Ral.

You Wu, David J Reiner.

  Ras is the most mutated oncoprotein in cancer. Among the three oncogenic effectors of Ras - Raf, PI3 Kinase and RalGEF>Ral - signalling through RalGEF>Ral (Ras-like) is by far the least well understood. A variety of signals and binding partners have been defined for Ral, yet we know little of how Ral functions in vivo. This review focuses on previous research in Drosophila that defined a function for Ral in apoptosis and established indirect relationships among Ral, the CNH-domain MAP4 Kinase misshapen, and the JNK MAP kinase basket. Most of the described signalling components are not essential in C. elegans, facilitating subsequent analysis using developmental patterning of the C. elegans vulval precursor cells (VPCs). The functions of two paralogous CNH-domain MAP4 Kinases were defined relative to Ras>Raf, Notch and Ras>RalGEF>Ral signalling in VPCs. MIG-15, the nematode ortholog of misshapen, antagonizes both the Ral-dependent and Ras>Raf-dependent developmental outcomes. In contrast, paralogous GCK-2, the C. elegans ortholog of Drosophila happyhour, propagates the 2°-promoting signal of Ral. Manipulations via CRISPR of Ral signalling through GCK-2 coupled with genetic epistasis delineated a Ras>RalGEF>Ral>Exo84>GCK-2>MAP3KMLK-1> p38PMK-1 cascade. Thus, genetic analysis using invertebrate experimental organisms defined a cascade from Ras to p38 MAP kinase.

Keywords:  Exo84; Exoc-8; Exocyst; LET-60; MLK-1; PMK-1; Ral-1; Rgl-1; Sec-5; Sec5; hppy; msn

DOI:  https://doi.org/10.1080/21541248.2021.1917953
Front Cell Dev Biol. 2021 ;9 631163

Taurine Attenuates the Hypotaurine-Induced Progression of CRC via ERK/RSK Signaling.

Xiaodan Hou, Junwei Hu, Xinyu Zhao, Qing Wei, Rongping Zhao, Min Li, Qiong Li.

  Colorectal cancer (CRC) is one of the most common malignant tumors, and previous metabolomics work has demonstrated great promise in identifying specific small molecules of tumor phenotype. In the present study, we analyzed the metabolites of resected tissues through gas chromatography-mass spectrometry (GC-MS), and found that the concentration of taurine in CRC tissues diminished whereas the concentration of hypotaurine increased. The results in vitro demonstrated that taurine significantly suppressed cellular proliferation, metastasis, and colony formation whereas it induced apoptosis in CRC cells. Furthermore, taurine regulated the expression levels of epithelial mesenchymal transition (EMT)-associated genes in a dose-dependent manner. Taurine also alleviated hypotaurine-induced CRC progression, which was linked to the inhibition of the ERK/RSK-signaling pathway and diminution in intracellular hypotaurine. Taurine additionally attenuated hypotaurine-induced tumor growth and metastasis in vivo. Patients with CRC exhibited lower levels of serum taurine, suggesting that taurine might be a promising biomarker reflecting a poor prognosis in CRC. Collectively, our results demonstrated that taurine-attenuated, hypotaurine-induced CRC progression provides a potential target for CRC therapy.

Keywords:  ERK; colorectal cancer; hypotaurine; prognosis; taurine; tumor progression

DOI:  https://doi.org/10.3389/fcell.2021.631163
Cancers (Basel). 2021 Apr 29. pii: 2148. [Epub ahead of print]13(9):

Discordance of KRAS Mutational Status between Primary Tumors and Liver Metastases in Colorectal Cancer: Impact on Long-Term Survival Following Radical Resection.

Francesco Ardito, Francesco Razionale, Lisa Salvatore, Tonia Cenci, Maria Vellone, Michele Basso, Elena Panettieri, Maria Alessandra Calegari, Giampaolo Tortora, Maurizio Martini, Felice Giuliante.

  If KRAS mutation status of primary colorectal tumor is representative of corresponding colorectal liver metastases (CRLM) mutational pattern, is controversial. Several studies have reported different rates of KRAS discordance, ranging from 4 to 32%. Aim of this study is to assess the incidence of discordance and its impact on overall survival (OS) in a homogenous group of patients. KRAS mutation status was evaluated in 107 patients resected for both primary colorectal tumor and corresponding CRLM at the same institution, between 2007 and 2018. Discordance rate was 15.9%. Its incidence varied according to the time interval between the two mutation analyses (p = 0.025; Pearson correlation = 0.2) and it was significantly higher during the first 6 months from the time of primary tumor evaluation. On multivariable analysis, type of discordance (wild-type in primary tumor, mutation in CRLM) was the strongest predictor of poor OS (p < 0.001). At multivariable logistic regression analysis, the number of CRLM >3 was an independent risk factor for the risk of KRAS discordance associated with the worst prognosis (OR = 4.600; p = 0.047). Results of our study suggested that, in the era of precision medicine, possibility of KRAS discordance should be taken into account within multidisciplinary management of patients with metastatic colorectal cancer.

Keywords:  KRAS discordance; KRAS mutation status; colorectal liver metastases; colorectal tumor; overall survival; precision medicine

DOI:  https://doi.org/10.3390/cancers13092148
Cell Signal. 2021 Apr 28. pii: S0898-6568(21)00121-2. [Epub ahead of print] 110032

Regulation of PCTAIRE1 protein stability by AKT1, LKB1, and BRCA1.

Syed Qaaifah Gillani, Misbah Un Nisa, Zarka Sarwar, Irfana Reshi, Sameer Bhat, Nusrat Nabi, Shaida Andrabi.

  PCTAIRE1, also known as CDK16, is a cyclin-dependent kinase that is regulated by cyclin Y. It is a member of the serine-threonine family of kinases and its functions have primarily been implicated in cellular processes like vesicular transport, neuronal growth and development, myogenesis, spermatogenesis and cell proliferation. However, as extensive studies on PCTAIRE1 have not yet been conducted, the signaling pathways for this kinase involved in governing many cellular processes are yet to be elucidated in detail. Here, we report the association of PCTAIRE1 with important cellular proteins involved in major cell signaling pathways, especially cell proliferation. In particular, here we show that PCTAIRE1 interacts with AKT1, a key player of the PI3K signaling pathway that is responsible for promoting cell survival and proliferation. Our studies show that PCTAIRE1 is a substrate of AKT1 that gets stabilized by it. Further, we show that PCTAIRE1 also interacts with and is degraded by LKB1, a kinase that is known to suppress cellular proliferation and also regulate cellular energy metabolism. Moreover, our results show that PCTAIRE1 is also degraded by BRCA1, a well-known tumor suppressor. Together, our studies highlight the regulation of PCTAIRE1 by key players of the major cell signaling pathways involved in regulating cell proliferation, and therefore, provide crucial links that could be explored further to elucidate the mechanistic role of PCTAIRE1 in cell proliferation and tumorigensis.

Keywords:  AKT1; BRCA1; Cell proliferation; LKB1; PCTAIRE1; PI3K

DOI:  https://doi.org/10.1016/j.cellsig.2021.110032
J Med Chem. 2021 May 07.

Breaking Oncogene Addiction: Getting RTK/RAS-Mutated Cancers off the SOS.

Erin Sheffels, Robert L Kortum.

In RTK/RAS-mutated cancers, therapeutic resistance is driven by rebound activation of multiple RTKs; broad inhibition of RTK signaling can potentially delay therapeutic resistance for a majority of patients. A new SOS1 inhibitor, BI-3406, broadly inhibits proximal RTK signaling will greatly expand the efficacy of therapies used to treat RTK/RAS-mutated cancers.

DOI: https://doi.org/10.1021/acs.jmedchem.1c00698
Sci Rep. 2021 May 04. 11(1): 9478

The genomic landscape of carcinomas with mucinous differentiation.

Bastien Nguyen, Francisco Sanchez-Vega, Christopher J Fong, Walid K Chatila, Amir Momeni Boroujeni, Fresia Pareja, Britta Weigelt, Christos Sotiriou, Denis Larsimont, Jorge S Reis-Filho, Christine Desmedt, Nikolaus Schultz.

Mucinous carcinomas can arise in any organ with epithelial cells that produce mucus. While mucinous tumors from different organs are histologically similar, it remains to be elucidated whether they share molecular alterations. Here we analyzed a total of 902 patients across six cancer types by comparing mucinous and non-mucinous samples, integrating text mining of pathology reports, gene expression, methylation, mutational and copy-number profiling. We found that, in addition to genes involved in mucin processing and secretion, MUC2 up-regulation is a multi-cancer biomarker of mucinous histology and is regulated by DNA methylation in colorectal, breast and stomach cancer. The majority of carcinomas with mucinous differentiation had fewer DNA copy-number alterations than non-mucinous tumors. The tumor mutational burden was lower in breast and lung with mucinous differentiation compared to their non-mucinous counterparts. We found several differences in the frequency of oncogenic gene and pathway alterations between mucinous and non-mucinous carcinomas, including a lower frequency of p53 pathway alterations in colorectal and lung cancer, and a lower frequency of PI-3-Kinase/Akt pathway alterations in breast and stomach cancer with mucinous differentiation. This study shows that carcinomas with mucinous differentiation originating from different organs share transcriptomic and genomic similarities. These results might pave the way for a more biologically relevant taxonomy for these rare cancers.

DOI: https://doi.org/10.1038/s41598-021-89099-2
Cell Rep. 2021 May 04. pii: S2211-1247(21)00400-9. [Epub ahead of print]35(5): 109069

mTOR signaling regulates gastric epithelial progenitor homeostasis and gastric tumorigenesis via MEK1-ERKs and BMP-Smad1 pathways.

Ke Li, Hongguang Wu, Ao Wang, Jean Charron, Yuji Mishina, Samy L Habib, Huijuan Liu, Baojie Li.

  mTOR, the sensor of nutrients and growth factors, has important roles in tissue homeostasis and tumorigenesis. However, how mTOR controls gastric epithelial cell turnover and gastric cancer development, a leading malignancy, remains poorly understood. Here, we provide genetic evidence that mTOR activation promotes proliferation and inhibits differentiation of Lgr5+ gastric epithelial progenitors (GEPs) in gastric homeostasis and tumorigenesis. mTOR signaling increases MEK1 and Smad1 expression and enhances activation of MEK1-ERKs and BMP-Smad1 pathways, respectively, in GEPs and gastric tumors. Mek1 deletion or inhibition rescues hyperproliferation, whereas Bmpr1a ablation or inhibition rescues differentiation defects of Tsc1-/- GEPs. Tsc1 deficiency in Lgr5+ GEPs accelerates gastric tumor initiation and development, which require MEK1-ERKs for hyperplasia and BMP-Smad1 for differentiation suppression. These findings reveal how mTOR signaling controls Lgr5+ GEP homeostasis and cancerization and suggest that ERKs and Smad1 signaling can be safely targeted to substitute mTOR inhibitors in gastric cancer therapy.

Keywords:  Lgr5; MEK1; Smad1; TSC; differentiation; gastric cancer; gastric epithelial progenitor; homeostasis; mTOR; tumorigenesis

DOI:  https://doi.org/10.1016/j.celrep.2021.109069