Dataset Information

Molecular characterization of perineural invasion-competent cancer cells using a novel in vitro model

ABSTRACT: Invasion of cancer cells into peripheral nerves, known as perineural invasion (PNI), is an established risk factor for cancer recurrence and metastasis. However, there are no approaches available to specifically target PNI-competent cancer cells for diagnostic and therapeutic purposes. The lack of ideal preclinical models simulating natural PNI challenges the true characterization of PNI cells and understanding of the cellular events underlying PNI. In this work, we developed an in vitro model recapitulating the biological events associated with natural PNI. The novel in vitro model included intact nerve preparations in 3D culture format to facilitate natural PNI. Using the novel model, we found that autonomic neurosignaling promotes PNI demonstrating the utility of this model in exploring the fundamental biology of PNI. Using this model, we also demonstrated that PNI cells have survival advantages against cancer chemotherapy further demonstrating the utility of this model in screening potential anti-PNI therapies. We also characterized the molecular profile of PNI-competent breast cancer cells purified from the in vitro model. Follow-up analysis showed that CPLANE1 and hemopexin are potential markers of PNI cells for future theranostics purposes. Overall, this work presents a new in vitro model of PNI, which can be used to study the fundamentals of PNI and screen potential anti-PNI therapies.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Breast Cancer

SUBMITTER: Nickson Joseph

LAB HEAD: Anand Krishnan

PROVIDER: PXD059147 | Pride | 2025-07-14

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Raw_Data.zip	Other
	SpectrumMill_DataSearch.zip	Other

Items per page:

1 - 2 of 2

Publications

Defining the cellular and molecular features of nerve-invaded cancer cells using a newly characterized experimental model.

Joseph Nickson N Shrestha Sanskriti S Sassmannshausen Ivanka I Mitzkus Sandra S Chumala Paulos P Sivakumar Bhadrapriya B Baiju Viswanath V Ahmed Shahid S Rees Henrike H Katselis George S GS Krishnan Anand A

Cell death discovery 20250708 1

Perineural invasion (PNI) is the invasion of cancer cells into nerves. Although PNI is a risk factor for cancer recurrence and metastasis, the lack of in vitro experimental models representing natural PNI challenges basic studies and therapeutic screening. In this work, we fully characterized a dorsal root ganglia (DRG)-nerve explant model for PNI and demonstrated the characteristic cellular and molecular features of cancer cells undergoing natural PNI. Briefly, thoracic and lumbar DRGs intactly ...[more]

PMID: 40628700

Similar Datasets

Project description:Aberrant activation of androgen receptor (AR)-dependent transcriptional programs is a hallmark of human prostate cancers. At the molecular level, ligand-mediated AR activation is coordinated through spatial and temporal protein-protein interactions (PPIs) involving AR-interacting proteins, which we designate the “AR-interactome”. Despite many years of research, the ligand-sensitive protein complexes involved in ligand-mediated AR activation in prostate-tumor cells has not been clearly defined. Here, we describe the development, characterization, and utilization of a novel human LNCaP prostate-tumor cell line, N-AR, which stably expresses wild-type AR containing the streptavidin-binding peptide epitope tagged at its N-terminus (SBP-AR). A bioanalytical workflow involving streptavidin-chromatography and label-free quantitative mass spectrometry was used to identify SBP-AR and associated ligand-sensitive proteins/protein complexes functionally linked to AR activation in the cytosol of N-AR cells. Functional studies verified that ligand-sensitive streptavidin-copurified proteins encoded modulators of AR-mediated transcription, suggesting that these novel proteins were putative SBP-AR-interacting proteins in N-AR cells. This was supported by biochemical associations between recombinant SBP-AR and the ligand-sensitive COPI retrograde trafficking complex in vitro. Extensive biochemical and molecular experiments showed that the COPI-retrograde complex regulates ligand-mediated AR transcriptional activation through the mobilization of Golgi-localized ARA160 coactivator into the nuclear compartment of prostate-tumor cells. Collectively, this study provides a bioanalytical strategy to validate the AR-interactome and define novel AR-interacting proteins involved in ligand-mediated AR activation in prostate-tumor cells. Moreover, we describe a cellular system to study how compartment-specific AR-interacting proteins influence AR activation and contribute to aberrant AR-dependent transcription that underlies the majority of human prostate cancers.

Project description:Fascioliasis is a major neglected tropical disease that infects humans and ruminant species worldwide. Recent discoveries have identified that liver fluke extracellular vesicles are released into the host’s internal environment. To understand extracellular vesicle biology further, isolation of the extracellular vesicle is necessary for downstream applications. The most widely used purification method involves differential centrifugation however; there is no ‘gold standard’ to date of extracellular vesicle isolation. Recently, size exclusion chromatography has been used to successfully purify EVs. In this study, liver flukes were collected to compare size exclusion chromatography EV purification, to the widely accepted method of differential centrifugation. Transmission electron microscopy and atomic force microscopy showed that size exclusion chromatography extracellular vesicles were smaller in size and diversity than differential centrifugation purified extracellular vesicles. Protein concentration and western blotting suggested that size exclusion chromatography purification had a high EV purity to protein yield ratio in comparison to differential centrifugation. Mass spectrometry analysis discovered a greater amount of protein identifications and unique peptide hits in differential centrifugation purified EVs with less diverse functionality, than size exclusion chromatography purified EVs. However, gene enrichment analysis against the Fasciola hepatica genome showed comparable results between purification methods. Furthermore, western blotting revealed that glutathione s-transferase sigma could be a potential marker for liver fluke EVs. These results suggest that size exclusion chromatography purification produces a higher EV purity to protein yield ratio and maintains extracellular vesicle composition, so therefore is a better suited method for further downstream experimentation with liver fluke extracellular vesicles, compared to differential centrifugation.