Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: Each box on the first row represents one of the 24 study participants (F508del/F508del CFTR, Aged 5-17 years) P.d; Participant de-identifier. Subsequent rows illustrate changes in treatment regimen, denoted by the switch in colour within columns. The colour indicates the specific CFTR modulators administered, including LUM/IVA (lumacaftor/ivacaftor), TEZ/IVA (tezacaftor/ivacaftor), or ETI (elexacaftor/tezacaftor/ivacaftor). B) Readouts. Clinical response (in vivo) : measured via lung function (FEV1pp) and sweat chloride (SC) pre- and post-treatment. Genetics: Comprehensive CFTR and pharmacogene sequencing. Human Nasal Epithelial Cell (HNEC) culture response (in vitro): assessed using electrophysiological measurements of CFTR function in differentiated-HNEC cultures derived from each participant. C) Clinical, Genetic and HNEC culture Relationships. Clinical outcomes, including FEV1 percent predicted and sweat chloride levels, are compared with genetic sequencing data and to differentiated-HNEC culture responses to CFTR modulators to evaluate the predictive value of epithelial cell models. FEV1pp: Forced Expiratory Volume in 1 second, percent predicted. Created in BioRender. Waters, S. (2025) https://BioRender.com/v19c483 457×139mm (118 × 118 DPI)

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques: In Vivo, Sequencing, In Vitro, Derivative Assay

Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: Waterfall plots illustrate the absolute change in participant outcomes after treatment with CFTR modulators. A) Change in FEV1pp. Absolute change in FEV1pp following treatment with lumacaftor/ivacaftor (LUM/IVA) or tezacaftor/ivacaftor (TEZ/IVA). Each bar represents an individual participant, and dotted lines indicate significant changes from baseline. B) Change in SC : Absolute change in SC levels after treatment with LUM/IVA or TEZ/IVA. Each bar represents a participant, with significant changes indicated by dotted lines. C) Subsequent Change in FEV1 : Absolute change in FEV1pp after switching directly to elexacaftor/tezacaftor/ivacaftor (ETI). Change is calculated as the difference between the subsequent baseline following treatment with a modulator, taken immediately prior to the new modulator, and post treatment values for the new regimen. D) Subsequent Change in SC: Absolute change in SC levels after switching directly to ETI, calculated as the difference between baseline (no treatment) and post-treatment values for the new regimen. E) Subsequent Change in FEV1 in participants who transitioned from LUM/IVA to TEZ/IVA before switching to ETI : Absolute change in FEV1pp after switching to a subsequent CFTR modulator TEZ/IVA or elexacaftor/tezacaftor/ivacaftor (ETI). Change is calculated as the difference between the subsequent baseline following treatment with a modulator, taken immediately prior to the new modulator, and post treatment values for the new regimen F) Subsequent Change in SC in participants who transitioned from LUM/IVA to TEZ/IVA before switching to ETI : Absolute change in SC levels after switching to TEZ/IVA or ETI, calculated as the difference between baseline (no treatment) and post-treatment values for the new regimen. Participants are identified consistently across all four graphs using a participant de-identifier. Nine participants had incomplete SC data ( Table S2 ). Symbols denote participants with a baseline FEV1pp below 90 (∘) or above 90 (∧). FEV1pp: Forced Expiratory Volume in 1 second, percent predicted. SC: Sweat Chloride. 182×220mm (300 × 300 DPI)

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques:

Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: Waterfall plots illustrate the absolute change in participant outcomes after treatment with CFTR modulators. Results are displayed with data grouped per participant. Participants are then grouped by the number of modulator treatments received (1, 2 or 3). Absolute change in FEV1pp following treatment with lumacaftor/ivacaftor (LUM/IVA), tezacaftor/ivacaftor (TEZ/IVA) or elexacaftor/ tezacaftor/ivacaftor (ETI) is displayed on the top graph with the corresponding participant’s absolute change in SC level displayed on the lower graph. Nine participants had incomplete SC data ( Table S2 ). Each bar represents an individual CFTR modulator response, with significant changes (FEV1pp change of >5 percentage points; SC decrease of > 20mmol/L) indicated by dotted lines. Participants are identified using a participant de-identifier. Symbols denote participants with a baseline FEV1pp below 90 (∘) or above 90 (∧). FEV1pp: Forced Expiratory Volume in 1 second, percent predicted. SC: Sweat Chloride.

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques:

Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: Pharmacogene function following treatment with A) Lumacaftor/Ivacaftor, B) Tezacaftor/Ivacaftor and C) Elexacaftor/Tezacaftor/Ivacaftor. The predicted activity levels of pharmacogenes were categorised for 58 genes from the Cytochrome P450 enzymes (CYP), ATP-binding cassette transporters (ABC), solute carrier families (SLC and SLCO) and UDP-glucuronosyltransferases (UGT). Activity levels are indicated based on their Stargazer score; increased (> 2), normal (= 2), intermediate (1 -2), poor (0 - 1) and unknown (< 0). Each column contains the predicted activity of a pharmacogene. Participants were ranked primarily by their responsiveness to treatment as measured by the relative increase in SC. FEV1pp was used to further rank participants with missing SC data. Each participant (n=24) is represented once per treatment and may be included up to 3 times. Pharmacogenes are not shown when the output for all participants was “normal” or “unknown”. SC: sweat chloride. FEV1pp: forced expiratory volume in one second, percent predicted.

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques: Activity Assay, Binding Assay

Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: A) Representative recordings and results from an individual participant. Changes in short-circuit current (ΔIsc) in differentiated human nasal epithelial cell (HNEC) culture from study participant number five. Cells were either untreated (0.01% DMSO vehicle) or pre-treated with a corrector(s) (Lumacaftor (LUM), tezacaftor (TEZ:T) or elexacaftor (E) plus TEZ for 48 hours). Functional CFTR expression was then measured by sequentially adding 100 μM apical amiloride (1. Amiloride), 0.01% apical DMSO vehicle control or 10 μM apical VX-770 (ivacaftor (IVA;I )), followed by 10 μM basal forskolin (3. Forskolin), 30 μM apical CFTR inhibitor (4. CFTRinh172), and 100 μM apical ATP (5. ATP). The basolateral-to-apical chloride gradient was used to measure functional CFTR activity. The i n vitro ΔIsc results for participant number five are displayed on the bar chart, with the dotted line indicating the wild-type reference level for our lab (as reported in Wong et al. 2022 42 ). CFTR activity is expressed as the inhibition of Isc by CFTRinh172 after activation by forskolin. Each dot represents an independent HNEC culture. Data are presented as mean ± SEM. B) ΔIsc in differentiated HNEC cultures to first modulator treatment. Bar chart showing ΔIsc (% of wild-type reference) in participants differentiated HNEC cultures in response to their first CFTR modulator treatment. CFTR activity is expressed as the inhibition of Isc by CFTRinh172 after activation by forskolin. Modulator response is calculated by subtracting baseline CFTR activity (DMSO) from modulator-treated results and reported as a percentage of normal (wild type). C) ΔIsc in differentiated HNEC cultures to subsequent modulator treatments. Bar chart showing ΔIsc (% of wild-type reference) in response to a subsequent CFTR modulator regimen. CFTR activity is expressed as the inhibition of Isc by CFTRinh172 after activation by forskolin. Changes are calculated by subtracting baseline CFTR activity (DMSO) from modulator-treated results and reported as a percentage of normal (wild type). Each bar represents the mean result of three replicate cultures per participant. Error bars represent the standard error of the mean (SEM). 113×233mm (300 × 300 DPI)

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques: Functional Assay, Expressing, Control, Activity Assay, Inhibition, Activation Assay

Journal: medRxiv

Article Title: INTEGRATING GENOMIC AND FUNCTIONAL TESTING TO IMPROVE CFTR MODULATOR RESPONSE PREDICTION IN CHILDREN WITH CYSTIC FIBROSIS

doi: 10.1101/2025.07.09.25331152

Figure Lengend Snippet: A) Patient Characteristics. The table summarises the characteristics of participants who switched to either tezacaftor/ivacaftor (TEZ/IVA) or elexacaftor/TEZ/IVA(ETI) treatments. Values for FEV1pp and SC are recorded immediately before switching treatments. Symbol (#) denotes three participants did not have measurements before switching to ETI. B) FEV1pp vs. CFTR activity (ΔIsc) in differentiated HNEC cultures. Plots of relative change in FEV1pp vs ΔIsc in response to CFTRinh172 treatment. Separate regression lines are shown for participants with baseline FEV1 above and below 90pp. The shaded area indicates the 95% confidence interval (CI). Each point (dot/triangle) represents a single comparison (n = 40). The colour of each point represents the specific CFTR modulator treatment administered C) SC vs. CFTR activity (ΔIsc) in differentiated-HNEC cultures. Plots of change in SC vs ΔIsc in response to CFTRinh172 treatment. The shaded area indicates the 95% confidence interval (CI). Each point represents a single comparison (n = 30 due to missing SC data for nine participants ( Table S2 )). The colour of each point represents the specific CFTR modulator treatment administered. FEV1pp: Forced Expiratory Volume in 1 second, percent predicted. SC: Sweat Chloride. 180×151mm (300 × 300 DPI)

Article Snippet: Briefly, differentiated-HNEC cultures were pre-incubated with CFTR correctors (3 μM LUM (VX-809; Selleckchem 1565 [ ]), 5 μM TEZ (VX-661; Selleckchem S7059 [ ], or a cocktail of 18 μM TEZ and 3 μM elexacaftor (E) (VX-445; Selleckchem S8851) [ ] for 48h prior to assessment of ion transport electrophysiology.

Techniques: Activity Assay, Comparison

Journal: Biomedicines

Article Title: Simultaneous Quantification of Ivacaftor, Tezacaftor, and Elexacaftor in Cystic Fibrosis Patients’ Plasma by a Novel LC–MS/MS Method

doi: 10.3390/biomedicines11020628

Figure Lengend Snippet: Chromatograms obtained: a calibrator at the LLOQ in plasma (panel 1.a, ivacaftor; panel 2.a, tezacafor, panel 3.a, elexacaftor); deuterated internal standards (panel 1.b, ivacaftor-d9, panel 2.b, tezacafor-d4, panel 3.b, elexacaftor-d3). RT, retention time. NL, normalized level.

Article Snippet: Ivacaftor (ref. I940600), tezacaftor (ref. T321510), and elexacaftor (ref. E201545) were purchased from Toronto Research Chemicals (North York, ON, Canada).

Techniques:

Journal: Biomedicines

Article Title: Simultaneous Quantification of Ivacaftor, Tezacaftor, and Elexacaftor in Cystic Fibrosis Patients’ Plasma by a Novel LC–MS/MS Method

doi: 10.3390/biomedicines11020628

Figure Lengend Snippet: The mean calibration curves in plasma (9-point calibration curve) of ivacaftor ( A ), tezacafor ( B ), and elexacaftor ( C ), ranging from 0.008 to 12 mg/L. Area Ratio, ratio between area standard and IS.

Article Snippet: Ivacaftor (ref. I940600), tezacaftor (ref. T321510), and elexacaftor (ref. E201545) were purchased from Toronto Research Chemicals (North York, ON, Canada).

Techniques:

Journal: Biomedicines

Article Title: Simultaneous Quantification of Ivacaftor, Tezacaftor, and Elexacaftor in Cystic Fibrosis Patients’ Plasma by a Novel LC–MS/MS Method

doi: 10.3390/biomedicines11020628

Figure Lengend Snippet: Results of intra-day and inter-day accuracy and reproducibility assays (n = 5). The quality controls’ concentrations were 0.008, 0.04, 0.41, 5 mg/L for LLOQ, QClow, QC medium, and QC high, respectively (CV% is the coefficient of variation percentage).

Article Snippet: Ivacaftor (ref. I940600), tezacaftor (ref. T321510), and elexacaftor (ref. E201545) were purchased from Toronto Research Chemicals (North York, ON, Canada).

Techniques:

Journal: Cureus

Article Title: The Impact of Elexacaftor/Ivacaftor/Tezacaftor on Cystic Fibrosis Patients Who Acquire COVID-19 Infection

doi: 10.7759/cureus.29276

Figure Lengend Snippet: Patients not on EIT therapy were almost three times more likely to experience respiratory failure after COVID-19 infection. EIT: elexacaftor, ivacaftor, and tezacaftor

Article Snippet: Both were also required to have a diagnosis of COVID-19 infection, identified in the TriNetX system by laboratory code 9088, signifying “presence of SARS coronavirus 2 and related RNA.” The inclusion criteria for the EIT-treated cohort also included medication code 2256951, indicating treatment with elexacaftor.

Techniques: Infection

Journal: Cureus

Article Title: The Impact of Elexacaftor/Ivacaftor/Tezacaftor on Cystic Fibrosis Patients Who Acquire COVID-19 Infection

doi: 10.7759/cureus.29276

Figure Lengend Snippet: A comparison of ventilator dependence between patients treated with EIT and those who were not. EIT: elexacaftor, ivacaftor, and tezacaftor

Article Snippet: Both were also required to have a diagnosis of COVID-19 infection, identified in the TriNetX system by laboratory code 9088, signifying “presence of SARS coronavirus 2 and related RNA.” The inclusion criteria for the EIT-treated cohort also included medication code 2256951, indicating treatment with elexacaftor.

Techniques: Comparison