Nemo-Like Kinase in Development and Diseases: Insights from Mouse Studies

May combine two claims.

NLK interacts with 14-3-3ζ (YWHAZ) to repress expression of E-cadherin by preventing dimerization of 14-3-3ζ and inhibiting migration of non-small cell lung cancer cells.

“Recently, it was found that NLK interacts with 14-3-3ζ (also known as YWHAZ) to repress the expression of E-cadherin by preventing dimerization of 14-3-3ζ and inhibiting the migration of non-small cell lung cancer cells [24].”

• cites [24] Chen, 2020, NLK interacts with 14-3-3ζ to restore the expression of E-cadherin, Oncol. Rep. sourceDisputed

  The Results report that NLK inhibits 14-3-3ζ dimerization and restores E-cadherin expression repressed by 14-3-3ζ in A549 cells—the opposite of the claim that NLK represses E-cadherin. The Abstract and Introduction likewise say NLK restores E-cadherin and inhibits migration. Although the Discussion's concluding sentence says “represses,” it conflicts with the paper's reported findings. (LLM, conf=0.90)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured

May combine two claims.

NLK expression in mouse delays SBMA disease progression but cannot completely prevent SBMA phenotype onset.

“Taken together, NLK expression in mouse delays disease progression but cannot completely prevent SBMA phenotype onset [51].”

• cites [51] Todd, 2015, Nemo-like kinase is a novel regulator of spinal and bulbar muscular atrophy, Elife sourceDisputed

  The Discussion says that a 50% reduction in NLK expression may delay disease progression but cannot completely prevent the full SBMA phenotype. The claim omits the reduction and attributes the delay to NLK expression itself; the Discussion instead says NLK normally promotes the disease condition. (LLM, conf=0.87)

  Checked by a reasoning model — may be wrong.

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Activation of NLK-induced phosphorylation of LEF1 and dissociation from HDAC1 facilitated LEF1-mediated target gene transcription.

“Furthermore, it was shown that activation of NLK-induced phosphorylation of LEF1 and dissociation from histone deacetylase 1 (HDAC1) facilitated LEF1-mediated target gene transcription [21].”

• cites [21] Ota, 2012, NLK positively regulates Wnt/β-catenin signalling by phosphorylating LEF1 in neural progenitor cells, EMBO J. sourceSupported

May combine two claims.

Phosphorylation of TCF4 by NLK prevents DNA binding of the β-catenin-TCF4 complex and limits β-catenin-mediated target gene transcription through TCF/LEF.

“This phosphorylation prevents DNA binding of the β-catenin-TCF4 complex and limits β-catenin-mediated target gene transcription through TCF/LEF [16,17].”

• cites [16] Ishitani, 1999, The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF, Nature sourceNot checked — no full text of the source
• cites [17] Ishitani, 2003, Regulation of Lymphoid Enhancer Factor 1/T-Cell Factor by Mitogen-Activated Protein Kinase-Related Nemo-Like Kinase-Dependent Phosphorylation in Wnt/β-Catenin Signaling, Mol. Cell. Biol. sourceSupported

May combine two claims.

Direct binding of NLK to STAT3 results in phosphorylation of STAT3 at Ser727, requiring activity of the TAK1-NLK cascade.

“Direct binding of NLK to STAT3 resulted in phosphorylation of STAT3 at Ser727, which requires the activity of the TAK1-NLK cascade [22,23].”

• cites [22] Ohkawara, 2004, Role of the TAK1-NLK-STAT3 pathway in TGF-beta-mediated mesoderm induction, Genes Dev. sourceSupported
• cites [23] Kojima, 2005, STAT3 regulates Nemo-like kinase by mediating its interaction with IL-6-stimulated TGFβ-activated kinase 1 for STAT3 Ser-727 phosphorylation, Proc. Natl. Acad. Sci. USA sourcePartly supported

  The Results show that STAT3 and NLK interact in a complex and that the TAK1–NLK pathway contributes to STAT3 Ser-727 phosphorylation in IL-6 signaling. The interaction experiments do not establish direct binding, and the paper reports that TGFβ-activated NLK did not phosphorylate STAT3 at Ser-727. The claim omits that signaling-specific qualification. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

In zebrafish, NLK signalling contributed to midbrain tectum development by promoting neural progenitor cell proliferation.

“In zebrafish, NLK signalling contributed to midbrain tectum development by promoting neural progenitor cell proliferation [21].”

• cites [21] Ota, 2012, NLK positively regulates Wnt/β-catenin signalling by phosphorylating LEF1 in neural progenitor cells, EMBO J. sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

NLK has an extended amino-terminal domain consisting of histidine, proline, alanine and glutamine.

“NLK also has an extended amino-terminal domain consisting of histidine, proline, alanine and glutamine [15].”

• cites [15] Brott, 1998, Nlk is a murine protein kinase related to Erk/MAP kinases and localized in the nucleus, Proc. Natl. Acad. Sci. USA sourcePartly supported

  The Isolation and Characterization of the Nlk cDNA section describes an extended amino-terminal region extremely rich in histidine, proline, glutamine, and alanine. It says these four amino acids account for more than 70% of the region, not that the region consists solely of them. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

NLK was originally identified in 1994 as the Drosophila gene nemo required for proper rotation of photoreceptor cells in the eye during ommatidia morphogenesis.

“was originally identified in 1994 as the Drosophila gene nemo required for proper rotation of photoreceptor cells in the eye during ommatidia morphogenesis [14].”

• cites [14] Choi, 1994, Rotation of photoreceptor clusters in the developing drosophila eye requires the nemo gene, Cell sourceSupported

May combine two claims.

NLK was initially shown to be involved in various developmental processes, such as promotion of mesoderm specification in sea urchins, mesoderm patterning, patterning of midbrain and forebrain in zebrafish, and oocyte maturation in Xenopus Laevis and in mouse testes.

“NLK was initially shown to be involved in various developmental processes, such as the promotion of mesoderm specification in sea urchins [32], mesoderm patterning, patterning of midbrain and forebrain in zebrafish [33], as well as oocyte maturation in Xenopus Laevis [34] and in mouse testes [35].”

• cites [32] Croce, 2006, Nemo-like kinase (NLK) acts downstream of Notch/Delta signalling to downregulate TCF during mesoderm induction in the sea urchin embryo, Development sourceSupported
• cites [33] Thorpe, 2004, Nemo-like kinase is an essential co-activator of Wnt signaling during early zebrafish development, Development sourceNot checked — no full text of the source
• cites [34] Ota, 2011, Possible Involvement of Nemo-like Kinase 1 in Xenopus Oocyte Maturation as a Kinase Responsible for Pumilio1, Pumilio2, and CPEB Phosphorylation, Biochemistry sourceNot checked — no full text of the source

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Foxp3 transcriptional activity was regulated by TAK1-NLK-mediated phosphorylation, and NLK activity was required to maintain homeostatic Treg cell numbers.

“In detail, Foxp3 transcriptional activity was regulated by TAK1-NLK-mediated phosphorylation, and NLK activity was required to maintain homeostatic Treg cell numbers.”

• cites [46] Fleskens, 2019, Nemo-like Kinase Drives Foxp3 Stability and Is Critical for Maintenance of Immune Tolerance by Regulatory T Cells, Cell Rep. sourcePartly supported

  The Discussion states that TAK1–NLK-mediated phosphorylation modulates Foxp3 transcriptional activity. In Results (“TREG Cell-Specific NLK Deficiency Results in Inflammation”; page unavailable), young NLK-deficient mice had comparable percentages of Foxp3+ Treg cells, while that population declined with age. The authors attribute the decline to reduced Foxp3 protein stability, not reduced Treg survival or proliferation, so the claim that NLK is required to maintain homeostatic Treg cell numbers overstates the finding. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

May combine two claims.

Activation of NLK can directly phosphorylate HDAC1, which in turn binds to LEF1 followed by repressed transcriptional activation of Wnt-target genes, including those supporting cell proliferation.

“It was shown that activation of NLK can directly phosphorylate HDAC1, which in turn binds to LEF1 followed by repressed transcriptional activation of Wnt-target genes, including those supporting cell proliferation [37].”

• cites [37] Masoumi, 2017, NLK-mediated phosphorylation of HDAC1 negatively regulates Wnt signaling, Mol. Biol. Cell sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing).

The Wnt signalling pathway is an important regulator in all developmental stages of T-cells.

“Furthermore, with regard to T-cell development, the Wnt signalling pathway has been described as an important regulator in all developmental stages of T-cells [42,43,44].”

• cites [42] Ma, 2012, β-catenin/TCF-1 pathway in T cell development and differentiation, J. Neuroimmune Pharmacol. Off. J. Soc. Neuroimmune Pharmacol. sourcePartly supported

  Section 3.2 describes Wnt signalling as indispensable for thymocyte development and says it regulates multiple processes at multiple stages. It does not say it regulates all stages; section 3.3 identifies DN2 proliferation as the earliest point at which its regulatory role is reported. (LLM, conf=0.85)

  Experimental — accuracy not yet measured
• cites [43] Clevers, 2002, WNT signaling and lymphocyte development, Cell sourceNot checked — no full text of the source
• cites [44] Coffer, 2018, The Role of WNT Signaling in Mature T Cells: T Cell Factor Is Coming Home, J. Immunol. sourceNot checked — no full text of the source

May combine two claims.

The lung phenotype in NLK-deficient mice is due to deletion of NLK that cannot phosphorylate LEF1, leading to elevated VEGF expression that causes aberrant proliferation of pulmonary epithelial and endothelial cells.

“This was due to the deletion of NLK that could not phosphorylated LEF1, which led to elevated levels of VEGF expression that caused aberrant proliferation of pulmonary epithelial and endothelial cells [36].”

• cites [36] Ke, 2016, Nemo-like kinase regulates the expression of vascular endothelial growth factor (VEGF) in alveolar epithelial cells, Sci. Rep. sourceSupported

• AssumptionRests on an assumption: The phenotypes observed in NLK-deficient mice are caused by the deletion of NLK rather than by the LacZ insertion or genetic background.

May combine two claims.

The interaction between HSP27 and NLK mediates nuclear localisation and protects cancer cells from apoptosis.

“The interaction between heat shock protein 27 (HSP27) and NLK mediated the nuclear localisation and protected the cancer cells from apoptosis [29].”

• cites [29] Shaw-Hallgren, G., Masoumi, K.C., Zarrizi, R., Hellman, U., Karlsson, P., Helou, K., and Massoumi, R. (2014). Association of Nuclear-Localized Nemo-Like Kinase with Heat-Shock Protein 27 Inhibits Apoptosis in Human Breast Cancer Cells. PLoS ONE, 9. sourceSupported

May combine two claims.

Most studies investigating immune and blood cells have found canonical Wnt signalling to be integral, affecting haematopoietic stem cell self-renewal, B-cell development in the bone marrow, dendritic cell maturation, and T-cell activation and migration.

“Most of the studies investigating immune and blood cells have found canonical Wnt signalling to be integral, since it has been shown to affect parts of haematopoietic stem cell self-renewal, B-cell development in the bone marrow, dendritic cell maturation and T-cell activation and migration [40,41].”

• cites [40] Staal, 2008, WNT signalling in the immune system: WNT is spreading its wings, Nat. Rev. Immunol. sourceNot checked — no full text of the source
• cites [41] Chae, 2018, Canonical and Non-Canonical Wnt Signaling in Immune Cells, Trends Immunol. sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

May combine two claims.

NLK has a neuroprotective role in HD, since deletion of NLK caused accelerated brain dysfunction.

“Overall, the authors were able to show that NLK has a neuroprotective role in HD, since the deletion of NLK caused accelerated brain dysfunction [52].”

• cites [52] Jiang, 2020, Nemo-like kinase reduces mutant huntingtin levels and mitigates Huntington’s disease, Hum. Mol. Genet. sourcePartly supported

  The Results (‘Genetic reduction of NLK exacerbates neuropathology in HD mice’) report that a roughly 50% reduction of NLK worsened brain atrophy and lowered DARPP32 in HD mice. The Discussion concludes that NLK has a protective role and that lower NLK levels may accelerate brain dysfunction. The study tested partial genetic reduction, not complete deletion, and hedges the acceleration claim. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

May combine two claims.

Inducing NLK in knock-out HD models resulted in attenuated brain atrophy and a reduction in mHTT aggregates.

“In addition, inducing NLK in the knock-out HD models resulted in attenuated brain atrophy and a reduction in mHTT aggregates.”

• cites [52] Jiang, 2020, Nemo-like kinase reduces mutant huntingtin levels and mitigates Huntington’s disease, Hum. Mol. Genet. sourcePartly supported

  The Results section reports that NLK overexpression in N171-82Q HD mice attenuated brain atrophy and reduced mHTT aggregates. But these were not knock-out HD models: the separate experiment involving NLK+/− mice reduced NLK and exacerbated brain atrophy. (LLM, conf=0.90)

  Experimental — accuracy not yet measured

May combine two claims.

Deletion of NLK could rescue muscle atrophy, motor neuron phenotypes, and extend the lifespan of heterozygous NLK mice compared to control mice in an SBMA model.

“Deletion of NLK could rescue the muscle atrophy, as well as motor neuron phenotypes, and furthermore extend the lifespan of the heterozygous NLK mice compared to control mice.”

• cites [51] Todd, 2015, Nemo-like kinase is a novel regulator of spinal and bulbar muscular atrophy, Elife sourceSupported

May combine two claims.

E15.5 NLK-deficient thymi were structurally normal and expressed normal Foxn1 levels.

“Histological examination of E15.5 NLK-deficient thymi revealed a structurally normal thymus that expressed normal Foxn1 levels, thereby suggesting that NLK is not required in the early stages of thymus development [48].”

• cites [48] Swann, 2017, Elevated levels of Wnt signaling disrupt thymus morphogenesis and function, Sci. Rep. sourceSupported

May combine two claims.

These cardiac-specific NLK transgenic mice developed baseline cardiomyopathy and, with pressure overload, were more susceptible to heart failure.

“These mice developed baseline cardiomyopathy and, with pressure overload, were more susceptible to heart failure.”

May combine two claims.

NLK-deficient mice had a significant reduction in single positive (SP) CD8+ thymocytes, without any effects on the SP CD4+ thymocyte population.

“Nonetheless, NLK-deficient mice had a significant reduction in single positive (SP) CD8+ thymocytes, without any effects on the SP CD4+ thymocyte population.”

• cites [45] Daams, 2020, Deletion of Nemo-like Kinase in T Cells Reduces Single-Positive CD8(+) Thymocyte Population, J. Immunol. sourceNot checked — no full text of the source

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MEF cells isolated from NLK-deficient animals show increased cell proliferation and shortened cell cycle compared to wild-type cells.

“Furthermore, using mouse embryonic fibroblast (MEF) cells isolated from NLK-deficient animals, we could show an increase in cell proliferation and shortened cell cycle compared to wild-type cells.”

May combine two claims.

Lungs of NLK-deficient mice show smaller and compressed alveoli and hyperthickening of the lung vasculature.

“The lungs of these mice showed smaller and compressed alveoli in addition to hyperthickening of the lung vasculature.”

May combine two claims.

Conditional deletion of NLK in the T-cell lineage, including early T-cell progenitor cells, did not affect the health, white and red blood cells, nor lymphoid tissue development of the mice.

“This deletion did not affect the health, white and red blood cells nor lymphoid tissue development of the mice.”

• cites [45] Daams, 2020, Deletion of Nemo-like Kinase in T Cells Reduces Single-Positive CD8(+) Thymocyte Population, J. Immunol. sourceNot checked — no full text of the source

May combine two claims.

NLK-deficient mice on a 129/Sv background survived up to 4-6 weeks after birth but displayed a pronounced cerebellar ataxia.

“NLK-deficient animals on a C57BL/6 background died in the third trimester of pregnancy, while mice on a 129/Sv background survived up to 4–6 weeks after birth, but displayed a pronounced cerebellar ataxia.”

• cites [47] Kortenjann, 2001, Abnormal bone marrow stroma in mice deficient for nemo-like kinase, Nlk, Eur. J. Immunol. sourceNot checked — no full text of the source

May combine two claims.

On a mixed background, NLK-deficient mice showed a severely compromised haematopoietic system, including a reduction in haematopoietic cells, fat tissue in the bone marrow cavity, and a loss of bone-lining cells.

“On a mixed background, NLK-deficient mice showed a severely compromised haematopoietic system, including a reduction in haematopoietic cells, fat tissue in the bone marrow cavity, as well as a loss of bone-lining cells.”

• cites [47] Kortenjann, 2001, Abnormal bone marrow stroma in mice deficient for nemo-like kinase, Nlk, Eur. J. Immunol. sourceNot checked — no full text of the source

May combine two claims.

In colorectal cancer, NLK inhibits cell growth and promotes cell apoptosis via a p53-independent signalling pathway.

“For example, in colorectal cancer, NLK inhibits cell growth and promotes cell apoptosis via a p53-independent signalling pathway [28].”

• cites [28] Yasuda, 2003, Nemo-like kinase induces apoptosis in DLD-1 human colon cancer cells, Biochem. Biophys. Res. Commun. sourceNot checked — no full text of the source

May combine two claims.

NLK is known as a negative regulator of Wnt signalling in most studies, but few studies show NLK positively promotes Wnt signalling.

“NLK is known as a negative regulator of Wnt signalling in most studies; however, there are few studies in which NLK has been shown to positively promote the Wnt signalling pathway [20].”

• cites [20] Ishitani, 2013, Nemo-like kinase, a multifaceted cell signaling regulator, Cell. Signal. sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

NARF, an E3 ubiquitin ligase, can form complexes with NLK.

“It has also been reported that NLK-associated RING finger protein (NARF), which is an E3 ubiquitin ligase, can form complexes with NLK.”

• cites [19] Yamada, 2006, NARF, an nemo-like kinase (NLK)-associated ring finger protein regulates the ubiquitylation and degradation of T cell factor/lymphoid enhancer factor (TCF/LEF), J. Biol. Chem. sourceNot checked — no full text of the source

May combine two claims.

Activation of NLK causes phosphorylation of TCF/LEF and ubiquitination-mediated degradation of this complex by NARF.

“Activation of NLK caused phosphorylation of TCF/LEF and ubiquitination-mediated degradation of this complex by NARF [19].”

• cites [19] Yamada, 2006, NARF, an nemo-like kinase (NLK)-associated ring finger protein regulates the ubiquitylation and degradation of T cell factor/lymphoid enhancer factor (TCF/LEF), J. Biol. Chem. sourceNot checked — no full text of the source

May combine two claims.

Dysregulation in NLK expression can affect the development and progression of various diseases, including cancer, where NLK acts as a tumour suppressor or oncogene depending on the tumour type.

“In addition, dysregulation in NLK expression can affect the development and progression of various diseases, including cancer, where NLK has been found to act as a tumour suppressor or oncogene depending on the tumour type [27].”

• cites [27] Huang, 2015, The emerging role of Nemo-like kinase (NLK) in the regulation of cancers, Tumor Biol. sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

NLK is an evolutionary conserved atypical proline-directed serine/threonine MAP kinase.

“Nemo-like kinase (NLK) is an evolutionary, conserved, atypical proline-directed serine/threonine mitogen-activated protein (MAP) kinase [13]”

• cites [13] Coulombe, 2007, Atypical mitogen-activated protein kinases: Structure, regulation and functions, Biochim. Biophys. Acta (BBA)-Mol. Cell Res. sourceNot checked — no full text of the source

May combine two claims.

The reduction in SP CD8+ thymocytes was caused by an increase in cell death and reduced phosphorylation of LEF1 and HDAC1.

“This reduction in SP CD8+ thymocytes was caused by an increase in cell death and reduced phosphorylation of LEF1 and HDAC1, suggesting that NLK plays an important role in the survival of CD8+ thymocytes [45].”

• cites [45] Daams, 2020, Deletion of Nemo-like Kinase in T Cells Reduces Single-Positive CD8(+) Thymocyte Population, J. Immunol. sourceNot checked — no full text of the source

May combine two claims.

NLK is expressed in biopsied tissues of melanoma patients, but expressed significantly lower in metastatic melanoma.

“NLK was found to be expressed in biopsied tissues of melanoma patients, but was expressed significantly lower in metastatic melanoma”

• cites [30] Yang, 2019, Decreased expression of nemo-like kinase in melanoma is correlated with increased vascularity and metastasis, Melanoma Res. sourceNot checked — no full text of the source

May combine two claims.

NLK acts as a master regulator for function and differentiation of cells from the immune system, including CD8+ SP thymocytes, Treg cells and regulation of the anti-viral response.

“Furthermore, NLK acts as a master regulator for function and differentiation of cells from the immune system, including CD8+ SP thymocytes, Treg cells and regulation of the anti-viral response (See Figure 1).”

Probably not a substantial claim: may not pass S4 (based). May combine two claims.

NLK was discovered as a negative regulator of the Wnt signalling pathway, but in later years it has been shown to be able to positively regulate Wnt signalling, depending on the organism and developmental processes.

“Traditionally, NLK was discovered as a negative regulator of the Wnt signalling pathway, but in later years it has been shown to be able to positively regulate Wnt signalling, depending on the organism and developmental processes.”