michela coan

What is the full name of the lncRNA renamed CHiLL1 in this study?

Cancer Hallmark in Lung LncRNA 1 (CHiLL1).

What type of cancer and mutation is the focus of this study?

KRAS-mutant non-small cell lung cancer (NSCLC).

How many lncRNAs were targeted in the CRISPR deletion library?

831 lncRNAs.

What CRISPR system was used for pooled deletion of lncRNA transcription start sites?

DECKO (dual-excision CRISPR knockout) CRISPR-deletion system.

What were the three cancer hallmarks screened in this study?

Proliferation, chemoresistance (cisplatin), and migration/invasion.

How many paired guide RNAs (pgRNAs) were designed per target?

10 unique pgRNAs per target.

What two KRAS+ NSCLC cell lines were used for parallel screening?

A549 and H460.

What method was used to validate that screen hits function via RNA rather than DNA elements?

Antisense oligonucleotide (ASO) knockdown.

How many high-confidence oncogenic lncRNAs were identified overall?

80.

What percentage of screened lncRNAs were identified as pan-hallmark hits?

Approximately 8%.

What is the subcellular localization of CHiLL1?

Principally cytoplasm.

What is the subcellular localization of GCAWKR?

Principally nucleus.

What pathway was enriched among CHiLL1-regulated genes related to cell death?

Apoptosis.

Knockdown of which lncRNA increased the percentage of cells in G2 phase?

GCAWKR.

What percentage of KRAS+ NSCLC tumors express at least one of CHiLL1 or GCAWKR?

78%.

What advantage did ASO cocktails show over single ASOs?

Greater effect on cell viability without increased toxicity in non-cancerous cells.

What is the name of the lncRNA that promotes gastric cancer and is also studied here?

GCAWKR.

What evidence suggested that lncRNA hits are clinically relevant?

They are amplified and overexpressed in tumors and associated with poor prognosis.

What method was used for 3D tumor model validation of ASOs?

Spheroid and patient-derived xenograft (PDX) organoid models.

What fraction of screen hits were successfully validated by ASOs as RNA-dependent?

8 out of 12 tested (approximately 67%).

What integration method was developed to combine multiple screen results?

Target Prioritisation Pipeline (TPP) using Robust Rank Aggregation and empirical Brown’s method.

What was the performance improvement of TPP over individual screens?

Higher AUC for correctly identifying positive and neutral controls.

What was the effect of NKAPL demethylation in the ovarian cancer paper, and how does it compare to CHiLL1/GCAWKR targeting?

NKAPL demethylation increased platinum sensitivity by 15%, similar to how ASOs against CHiLL1/GCAWKR reduced viability in NSC