Research ArticleImmunology Open Access | 10.1172/jci.insight.172312

Christina Michalski,1,2 Claire Cheung,1,2 Ju Hee Oh,1,3 Emma Ackermann,1 Constantin R. Popescu,1,2,4 Anne-Sophie Archambault,1,3 Martin A. Prusinkiewicz,1,2 Rachel Da Silva,1 Abdelilah Majdoubi,1,2 Marina Viñeta Paramo,1,5 Rui Yang Xu,1,5 Frederic Reicherz,1,2 Annette E. Patterson,1,3 Liam Golding,1,2,5 Ashish A. Sharma,6 Chinten J. Lim,1,2 Paul C. Orban,1,7 Ramon I. Klein Geltink,1,3 and Pascal M. Lavoie1,2,5

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

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1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Golding, L. in: JCI | PubMed | Google Scholar

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Sharma, A. in: JCI | PubMed | Google Scholar |

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Lim, C. in: JCI | PubMed | Google Scholar

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Orban, P. in: JCI | PubMed | Google Scholar

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Klein Geltink, R. in: JCI | PubMed | Google Scholar

1British Columbia Children’s Hospital Research Institute, Vancouver, British Columbia, Canada.

2Department of Pediatrics and

3Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

4Department of Pediatrics, Université Laval, Quebec, Quebec, Canada.

5Women+ and Children′s Health, Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.

6Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia, USA.

7Department of Surgery, University of British Columbia, Vancouver, British Columbia, Canada.

Address correspondence to: Pascal M. Lavoie, BC Children’s Hospital Research Institute, 5th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: plavoie@bcchr.ca. Or to: Ramon I. Klein Geltink, BC Children’s Hospital Research Institute, 4th Floor, Translational Research Building, 950 West 28th Avenue, Vancouver, British Columbia V5Z 4H4, Canada. Email: ramon.kleingeltink@bcchr.ca.

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Find articles by Lavoie, P. in: JCI | PubMed | Google Scholar

Authorship note: RIKG and PML are co–senior and co–corresponding authors.

Published February 6, 2024 - More info

Published in Volume 9, Issue 5 on March 8, 2024
JCI Insight. 2024;9(5):e172312. https://doi.org/10.1172/jci.insight.172312.
© 2024 Michalski et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published February 6, 2024 - Version history
Received: May 15, 2023; Accepted: January 31, 2024 View PDF Abstract

Pattern recognition receptor responses are profoundly attenuated before the third trimester of gestation in the relatively low-oxygen human fetal environment. However, the mechanisms regulating these responses are uncharacterized. Herein, genome-wide transcription and functional metabolic experiments in primary neonatal monocytes linked the negative mTOR regulator DDIT4L to metabolic stress, cellular bioenergetics, and innate immune activity. Using genetically engineered monocytic U937 cells, we confirmed that DDIT4L overexpression altered mitochondrial dynamics, suppressing their activity, and blunted LPS-induced cytokine responses. We also showed that monocyte mitochondrial function is more restrictive in earlier gestation, resembling the phenotype of DDIT4L-overexpressing U937 cells. Gene expression analyses in neonatal granulocytes and lung macrophages in preterm infants confirmed upregulation of the DDIT4L gene in the early postnatal period and also suggested a potential protective role against inflammation-associated chronic neonatal lung disease. Taken together, these data show that DDIT4L regulates mitochondrial activity and provide what we believe to be the first direct evidence for its potential role supressing innate immune activity in myeloid cells during development.

Introduction

The fetal immune system has a remarkable ability to tolerate maternal semiallogeneic antigens, in part through lowered innate immune activity, the presence of prominent antiinflammatory responses, and suppression from specialized, regulatory T and B cells (1). Pattern recognition receptor (PRR) responses are broadly suppressed in the fetus in early gestation, but the mechanisms involved are unclear (2, 3). Specifically, these responses become activated in the early third trimester of gestation in humans, following a hierarchical emergence pattern, with cytosolic/intracellular PRR showing activity arising before extracellular PRR in neonatal monocytes (4–6). These developmental changes are coordinated, such that, at the term of gestation, neonatal myeloid cells are able to produce robust responses that prepare the newborn for de novo exposure to microorganisms (7, 8). Although these functional changes have been well described, the molecular mechanisms underlying the regulation of innate immune responses early on, during human development, remain poorly understood.

Cellular metabolism plays an important role in regulating innate immune responses in general and, specifically also, during early ontogeny (2, 9). During homeostasis, immune cells engage energetically efficient oxidative phosphorylation in the mitochondria for ATP production. Robust inflammatory responses following PRR stimulation require rapid energy production, with upregulation of biosynthesis pathways through cytoplasmic glucose metabolism; conversely, alternative activation is regulated through mitochondrial pathways (10). When oxygen supply becomes limited, significant changes in metabolic pathway activity occur, with increased reliance on glycolysis and remodeling of the mitochondrial network by altering mitochondrial dynamics (11, 12). In utero, oxygen saturation is physiologically low (13), which raises the possibility that distinct cell-intrinsic mechanisms may be required to adapt to the metabolic conditions of the fetal environment. While physiological hypoxia maintains (hematopoietic) stem cells in a quiescent state (14), pathological hypoxia can also suppress homeostatic monocyte functions (15). mTORC1 is a key signaling hub for maintaining hematopoietic stem cell quiescence and stemness through limitation of mitochondrial biogenesis (16). mTORC1 activity is reduced in neonatal monocytes and monocyte-derived macrophages (2, 9). Additionally, our previous data indicated a potential role for DDIT4L in this context, as its expression was found to be increased in neonatal monocytes in unbiased genome-wide gene expression experiments (2).

DDIT4L, a paralog of DDIT4, negatively regulates mTORC1 (17, 18), which, in turn, is a key regulator of downstream processes such as proliferation, protein translation, and cell-intrinsic metabolism (19). In mice, targeted deletion of the Ddit4 gene reprograms bone marrow–derived macrophages toward a glycolytic phenotype and enhances LPS-induced IL-1β responses (20). In cardiomyocytes, diverse metabolic stress conditions, such as glucose or serum deprivation, cellular oxidative stress, or DNA damage, caused upregulation of DDIT4L (21). These data suggest that DDIT4L may play an important role in modulating cellular metabolic stress responses and innate immune activity during development.

Here, we tested whether DDIT4L can directly regulate innate immune responsiveness in human neonatal myeloid cells. Using a combination of molecular, functional, and metabolic studies, comparing primary preterm, term, and adult monocytes, we demonstrate a gestational age–dependent reduction in mitochondrial capacity in neonatal monocytes. Furthermore, we demonstrate that DDIT4L is selectively upregulated under metabolic stress conditions in neonatal monocytes. To provide more direct mechanistic insights into the potential role of DDIT4L in this context, we created genetically modified monocytic U937 cell clones. Biochemical, metabolic, and imaging studies using these cells confirmed that DDIT4L upregulation reduced mitochondrial mass, altered regulators of mitochondrial dynamics, and blunted LPS cytokine responses. Further analysis showed that DDIT4L is upregulated in lung macrophages from premature neonates during the neonatal period, with reduced expression of this gene in infants who develop bronchopulmonary dysplasia, suggesting a potential protective role against inflammatory-associated lung injury. Altogether, these data point toward a developmental role for DDIT4L in regulation of mitochondrial and innate immune activity in myeloid cells as well as adaption to metabolic stress and prevention of pathological inflammation during the fetal and early neonatal transition period.

Results

Oxidative metabolism and mitochondrial activity are reduced in neonatal monocytes. Neonatal monocytes and monocyte-derived macrophages displayed reduced mTOR and glycolytic activity at lower gestational age (2, 9). At baseline, oxygen consumption rates (OCRs) were comparable in preterm, full-term neonatal, and adult monocytes (Figure 1A). However, when normalized to mitochondrial mass, as measured by MitoTracker Green, neonatal monocytes showed significantly less respiration (Figure 1, B and C). Additionally, spare respiratory capacity (SRC) was further reduced at lower gestational age, as shown by a profound reduction of respiration in response to carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, an ionophore that uncouples mitochondrial oxidative phosphorylation from ATP production), specifically in preterm neonatal monocytes, compared with term or adult monocytes (Figure 1, D and E). Normalization of SRC [calculated as (OCR after FCCP – basal OCR)/basal OCR × 100] to mitochondrial volume revealed that adult monocyte mitochondria were considerably more efficient than their neonatal counterparts, implying that higher mitochondrial mass may partly compensate for a reduced mitochondrial efficiency at term (Supplemental Figure 1; supplemental material available online with this article; https://doi.org/10.1172/jci.insight.172312DS1), but this efficiency was not seen earlier in gestation in preterm monocytes (Figure 1F). These data revealed a developmental gradient in mitochondrial respiratory capacity in myeloid cells.

Figure 1

Reduced mitochondrial activity and capacity in neonatal monocytes. (A) Mitochondrial activity (basal oxygen consumption rate [OCR]), (B) mitochondrial mass (Mitotracker Green staining by flow cytometry; mean fluorescent intensity [MFI]), and (C) ratio of basal OCR to mitochondrial mass (when sufficient samples were available for paired analyses) in adult, term, and preterm monocytes. (D) Mitochondrial stress test (mean with 95% confidence interval and with x axis values nudged by 1 minute between age groups to enhance visibility), with (E) spare respiratory capacity (SRC) and (F) the ratio of SRC to mitochondrial mass (when sufficient samples were available for paired analyses). Data are from 15 adult, 13 term, and 6 preterm samples. (G) Nonmitochondrial oxygen consumption from same samples as in A, D, and E. (H) Expression of ROS genes (Hallmark pathway), using data from GEO accession GSE104510 (2). Data from 11 (LPS) to 12 (unstimulated) adults, 12 term and 6 (LPS) to 8 (unstimulated) preterm samples. (I) Electron transport chain proteins (by Western blot; data are from 5 adult, term, and preterm neonatal samples), including (J) a representative blot (of 5) of regulators and indicators of mitochondrial fission and fusion (K) TOM20, (L) DRP1, (M) DRP1S616 phosphorylation, (N) MFF, and (O) MFN2 in primary monocytes (see Supplemental Figure 2B for raw Western blot images). Data are from 5 adult, 5 term, and 4 preterm neonatal samples. For A–G, I, and K–O, data are represented as boxes (25th to 75th percentile) and whiskers (minimum to maximum), with medians indicated by solid lines. Groups were compared using 1-way ANOVA with Tukey’s multiple comparisons tests between age groups. For I, groups were compared using 2-way ANOVA, with nonsignificant age effect (P > 0.05). *P < 0.05, ***P < 0.001; only significant P values are shown.

Consistent with gestational age–dependent changes in mitochondrial activity in primary human neonatal monocytes, preterm neonatal monocytes also showed increased nonmitochondrial oxygen consumption (Figure 1G) and reduced expression of antioxidant genes, such as microsomal glutathione S-transferase 1 (MGST1), peroxiredoxin 4 (PRDX4), and superoxide dismutase (SOD1) (Figure 1H). These differences may indicate that preterm monocytes may be less resilient to hyperoxia. Furthermore, there were no differences in expression of mitochondrial electron transport chain complex proteins (Figure 1, I and J), supporting that the reduced mitochondrial reserve capacity in neonatal monocytes may be more related to altered mitochondrial morphology, as the former was previously shown to correlate with altered mitochondrial morphology in primary T cells (22). Indeed, when examining expression profiles of genes involved in mitochondrial fusion, gestational age–dependent developmental differences were observed, with preterm monocytes standing out, compared with term and adult monocytes (Supplemental Figure 2A).

The reduced mitochondrial activity and respiratory capacity in neonatal monocytes was also supported by significantly reduced levels of translocase of the outer mitochondrial membrane 20 (TOM20) in preterm monocytes, a proxy often used for mitochondrial mass (Figure 1K and Supplemental Figure 2B). Furthermore, total dynamin-related protein 1 (DRP1) was significantly reduced in preterm monocytes compared with term monocytes (Figure 1L), although we could not reproducibly detect the fission-associated phosphorylated form of DRP1ser616 (Figure 1M). Expression of the mitochondrial fission factor (MFF) was also reduced in preterm monocytes, and this was especially evident in the high-molecular-weight splice isoforms, although these differences were not significant, possibly due to the small number of sample replicates (Figure 1, M and N, and Supplemental Figure 2B). Expression of the mitochondrial fusion protein mitofusin-2 (MFN2) did not differ between the age groups (Figure 1O). Overall, our data could suggest that lower gestational age is associated with altered mitochondrial dynamics and reduced mitochondrial efficiency (22, 23). Yet, these data also support a higher propensity for ROS generation and oxidative stress in early gestation monocytes (24) in the context of reduced mitochondrial efficiency.

Metabolic limitations reduce cytokine production and induce DDIT4L expression. Next, we sought to determine whether