Summary of Study ST004001

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002506. The data can be accessed directly via it's Project DOI: 10.21228/M8QG1M This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST004001
Study Title	Metabolic and Lipidomic Trade-offs in Helicoverpa armigera: Dynamics Under Plant Protease Inhibitor-Induced Stress
Study Type	Metabolomics
Study Summary	Plant protease inhibitors retard the growth and development of insects by inhibiting their digestive proteases. In response, insects try to adapt to these plant defensive molecules by modulating their protease expression. However, their survival mechanisms might not be limited only to digestive plasticity. To explore this, we performed a comprehensive lipidomics and metabolomics analysis in Helicoverpa armigera fed with a recombinant Capsicum annuum protease inhibitor (rCanPI-7) having unique four inhibitory repeat domains with potent activity against insect trypsins and chymotrypsins. These results revealed that H. armigera employs a dynamic and multifaceted physiological response to dietary stress induced by rCanPI. Upon ingestion of rCanPI-7, down regulation of glycolysis and TCA cycle indicated a decrease in primary energy metabolism while oxidative stress was evident from the depletion of reduced glutathione, peroxidation of membrane lipids, and accumulation of ceramides which are the hallmarks of mitochondrial dysfunction. Investigation of the dynamics in the turnover of different molecules hints that H. armigera activated multiple compensatory strategies such as mobilizing triglycerides and amino acid catabolism as an alternative source of energy, upregulation of antioxidants, membrane remodeling, activation of apoptosis, and shifts in neuromodulatory metabolites linked to cognitive adaptation. Collectively, these findings point to a tightly regulated physiological tug-of-war in H. armigera, where the damaging impact of rCanPI-induced oxidative and nutritional stress is counteracted by a suite of compensatory metabolic, structural, and neuromodulatory adjustments. To our knowledge, this is the first report of lipidomic profiling in H. armigera, providing novel insights into its biochemical resilience and identifying potential metabolic vulnerabilities for enhancing biopesticide strategies.
Institute	Translational Health Science And Technology Institute (THSTI)
Department	NCD
Laboratory	Biomarker lab
Last Name	Kumar
First Name	Yashwant
Address	NCR Biotech Science Cluster,, Faridabad, Haryana, 121001, India
Email	y.kumar@thsti.res.in
Phone	+911292876796
Submit Date	2025-06-20
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2025-07-09
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002506
Project DOI:	doi: 10.21228/M8QG1M
Project Title:	Metabolic and Lipidomic Trade-offs in Helicoverpa armigera: Dynamics Under Plant Protease Inhibitor-Induced Stress
Project Type:	Metabolomics
Project Summary:	Plant protease inhibitors retard the growth and development of insects by inhibiting their digestive proteases. In response, insects try to adapt to these plant defensive molecules by modulating their protease expression. However, their survival mechanisms might not be limited only to digestive plasticity. To explore this, we performed a comprehensive lipidomics and metabolomics analysis in Helicoverpa armigera fed with a recombinant Capsicum annuum protease inhibitor (rCanPI-7) having unique four inhibitory repeat domains with potent activity against insect trypsins and chymotrypsins. These results revealed that H. armigera employs a dynamic and multifaceted physiological response to dietary stress induced by rCanPI. Upon ingestion of rCanPI-7, down regulation of glycolysis and TCA cycle indicated a decrease in primary energy metabolism while oxidative stress was evident from the depletion of reduced glutathione, peroxidation of membrane lipids, and accumulation of ceramides which are the hallmarks of mitochondrial dysfunction. Investigation of the dynamics in the turnover of different molecules hints that H. armigera activated multiple compensatory strategies such as mobilizing triglycerides and amino acid catabolism as an alternative source of energy, upregulation of antioxidants, membrane remodeling, activation of apoptosis, and shifts in neuromodulatory metabolites linked to cognitive adaptation. Collectively, these findings point to a tightly regulated physiological tug-of-war in H. armigera, where the damaging impact of rCanPI-induced oxidative and nutritional stress is counteracted by a suite of compensatory metabolic, structural, and neuromodulatory adjustments. To our knowledge, this is the first report of lipidomic profiling in H. armigera, providing novel insights into its biochemical resilience and identifying potential metabolic vulnerabilities for enhancing biopesticide strategies.
Institute:	Translational Health Science And Technology Institute (THSTI)
Department:	NCD
Laboratory:	Biomarker lab
Last Name:	Kumar
First Name:	Yashwant
Address:	NCR Biotech Science Cluster,, Faridabad, Haryana, 121001, India
Email:	y.kumar@thsti.res.in
Phone:	01292876796

Subject:

Subject ID:	SU004139
Subject Type:	Insect
Subject Species:	Helicoverpa armigera ( 29058)
Taxonomy ID:	29058

Factors:

Subject type: Insect; Subject species: Helicoverpa armigera ( 29058) (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Treatment
SA461366	EC_2	Early	Control
SA461367	EC_1	Early	Control
SA461368	EC_3	Early	Control
SA461369	EI_1	Early	Fed
SA461370	EI_2	Early	Fed
SA461371	EI_3	Early	Fed
SA461372	LC_1	Late	Control
SA461373	LC_2	Late	Control
SA461374	LC_3	Late	Control
SA461375	LI_2	Late	Fed
SA461376	LI_3	Late	Fed
SA461377	LI_1	Late	Fed
SA461378	MC_1	Mid	Control
SA461379	MC_2	Mid	Control
SA461380	MC_3	Mid	Control
SA461381	MI_1	Mid	Fed
SA461382	MI_2	Mid	Fed
SA461383	MI_3	Mid	Fed

Showing results 1 to 18 of 18

Showing results 1 to 18 of 18

Collection:

Collection ID:	CO004132
Collection Summary:	Three biological replicates of AD-fed and rCanPI-fed insects from early, mid, and late responses were used. Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μl was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μl of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C.
Sample Type:	Larvae

Treatment:

Treatment ID:	TR004148
Treatment Summary:	Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study.

Treatment ID:

TR004148

Treatment Summary:

Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study.

Sample Preparation:

Sampleprep ID:	SP004145
Sampleprep Summary:	Three biological replicates of AD-fed and rCanPI-fed insects from early, mid, and late responses were used. Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C.

Sampleprep ID:

SP004145

Sampleprep Summary:

Three biological replicates of AD-fed and rCanPI-fed insects from early, mid, and late responses were used. Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C.

Chromatography:

Chromatography ID:	CH005008
Chromatography Summary:	Metabolite separation was performed using an Ultimate 3000 UPLC system, employing both reverse-phase and hydrophilic interaction liquid chromatography columns under positive and negative ionization modes. HSS T3 column
Instrument Name:	Thermo Dionex Ultimate 3000 RS
Column Name:	Waters ACQUITY UPLC HSS T3 (100 x 2.1 mm, 1.8 μm)
Column Temperature:	40°C
Flow Gradient:	1% B to 95% B in 10 minutes
Flow Rate:	300 μL/min
Solvent A:	100% Water; 0.1% Formic acid
Solvent B:	100% Methanol; 0.1% Formic acid
Chromatography Type:	Reversed phase

Chromatography ID:	CH005009
Chromatography Summary:	XBridge BEH Amide (Waters Corporation, Milford, MA, USA) column were used for reverse-phase and HILIC, respectively. For polar compound separation, solvent A consisted of 20 mM ammonium acetate in water (pH 9.0), while mobile phase B was 100% acetonitrile.
Instrument Name:	Thermo Dionex Ultimate 3000 RS
Column Name:	Waters XBridge BEH Amide (100 x 2.1 mm, 2.5 μm)
Column Temperature:	40°C
Flow Gradient:	85% B and proceeds to 10% B over 14 minutes
Flow Rate:	200 μL/min
Solvent A:	100% Water; 20 mM ammonium acetate (pH-9.0)
Solvent B:	100% Acetonitrile
Chromatography Type:	HILIC

Analysis:

Analysis ID:	AN006596
Analysis Type:	MS
Chromatography ID:	CH005008
Has Mz:	1
Has Rt:	1
Rt Units:	Seconds
Results File:	ST004001_AN006596_Results.txt
Units:	relative intensity

Analysis ID:	AN006597
Analysis Type:	MS
Chromatography ID:	CH005008
Has Mz:	1
Has Rt:	1
Rt Units:	Seconds
Results File:	ST004001_AN006597_Results.txt
Units:	relative intensity

Analysis ID:	AN006598
Analysis Type:	MS
Chromatography ID:	CH005009
Has Mz:	1
Has Rt:	1
Rt Units:	Seconds
Results File:	ST004001_AN006598_Results.txt
Units:	relative intensity

Analysis ID:	AN006599
Analysis Type:	MS
Chromatography ID:	CH005009
Has Mz:	1
Has Rt:	1
Rt Units:	Seconds
Results File:	ST004001_AN006599_Results.txt
Units:	relative intensity