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Article ; Online: Generating Genetic Mosaic Mouse Embryos or Organoids for Studies of Kidney Development.

Costantini, Frank

Methods in molecular biology (Clifton, N.J.)

2019 Volume 1926, Page(s) 3–21

Abstract: For studies of gene function during development, it can be very useful to generate mosaic embryos in which a small subset of cells in a given cell lineage lacks a gene of interest and carries a marker that allows the mutant cells to be specifically ... ...

Abstract	For studies of gene function during development, it can be very useful to generate mosaic embryos in which a small subset of cells in a given cell lineage lacks a gene of interest and carries a marker that allows the mutant cells to be specifically visualized and compared to wild-type cells. Several methods have been used to generate genetically mosaic mouse kidneys for such studies. These include (1) chimeric embryos generated using embryonic stem cells, (2) chimeric renal organoids generated by dissociation and reaggregation of the fetal kidneys, (3) generation of a knockout allele with a built-in reporter gene, (4) mosaic analysis with double markers (MADM), and (5) mosaic mutant analysis with spatial and temporal control of recombination (MASTR). In this chapter, these five methods are described, and their advantages and disadvantages are discussed.
MeSH term(s)	Animals ; DNA Nucleotidyltransferases/genetics ; DNA Nucleotidyltransferases/metabolism ; Embryo, Mammalian/cytology ; Embryo, Mammalian/metabolism ; Embryonic Stem Cells/cytology ; Embryonic Stem Cells/metabolism ; Integrases/genetics ; Integrases/metabolism ; Kidney/cytology ; Kidney/embryology ; Kidney/metabolism ; Mice ; Mice, Knockout ; Organoids/cytology ; Organoids/metabolism
Chemical Substances	Cre recombinase (EC 2.7.7.-) ; DNA Nucleotidyltransferases (EC 2.7.7.-) ; FLP recombinase (EC 2.7.7.-) ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2019-02-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-9021-4_1
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Article ; Online: Ret signaling in ureteric bud epithelial cells controls cell movements, cell clustering and bud formation.

Packard, Adam / Klein, William H / Costantini, Frank

Development (Cambridge, England)

2021 Volume 148, Issue 9

Abstract: Ret signaling promotes branching morphogenesis during kidney development, but the underlying cellular mechanisms remain unclear. While Ret-expressing progenitor cells proliferate at the ureteric bud tips, some of these cells exit the tips to generate the ...

Abstract	Ret signaling promotes branching morphogenesis during kidney development, but the underlying cellular mechanisms remain unclear. While Ret-expressing progenitor cells proliferate at the ureteric bud tips, some of these cells exit the tips to generate the elongating collecting ducts, and in the process turn off Ret. Genetic ablation of Ret in tip cells promotes their exit, suggesting that Ret is required for cell rearrangements that maintain the tip compartments. Here, we examine the behaviors of ureteric bud cells that are genetically forced to maintain Ret expression. These cells move to the nascent tips, and remain there during many cycles of branching; this tip-seeking behavior may require positional signals from the mesenchyme, as it occurs in whole kidneys but not in epithelial ureteric bud organoids. In organoids, cells forced to express Ret display a striking self-organizing behavior, attracting each other to form dense clusters within the epithelium, which then evaginate to form new buds. The ability of forced Ret expression to promote these events suggests that similar Ret-dependent cell behaviors play an important role in normal branching morphogenesis.
MeSH term(s)	Animals ; Cell Movement ; Cluster Analysis ; Epithelial Cells/metabolism ; Epithelium/metabolism ; Female ; Kidney/growth & development ; Kidney/metabolism ; Male ; Mesoderm/metabolism ; Mice ; Mice, Inbred C57BL ; Morphogenesis ; Organoids ; Protein-Tyrosine Kinases/metabolism ; Signal Transduction ; Stem Cells/metabolism ; Ureter/metabolism
Chemical Substances	Protein-Tyrosine Kinases (EC 2.7.10.1)
Language	English
Publishing date	2021-04-29
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.199386
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Book ; Conference proceedings: Genetic manipulation of the early mammalian embryo

Costantini, Frank

[collected papers of the fall 1984 Banbury Conf. on the Genetic Manipulation of the Mammalian Ovum and Early Embryo, Lloyd Harbor, NY]

(Banbury report ; 20)

1985

Event/congress	Conference on the Genetic Manipulation of the Mammalian Ovum and Early Embryo (1984, LloydHarborNY)
Author's details	ed. by Frank Costantini
Series title	Banbury report ; 20
Collection
Keywords	Genetic Engineering ; Mammals / embryology ; Mammals / genetics ; Gentechnologie ; Embryo ; Säugetiere
Subject	Embryonen ; Mammalia ; Säuger ; Säugetier ; Gentechnik ; Genchirurgie ; Genetic engineering ; Genetische Manipulation ; Genetische Technik ; Genmanipulation
Size	XI, 289 S. : Ill., graph. Darst.
Publisher	Cold Spring Harbor Laboratory
Publishing place	Cold Spring Harbor
Publishing country	United States
Document type	Book ; Conference proceedings
HBZ-ID	HT002482829
ISBN	0-87969-220-0 ; 978-0-87969-220-9
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Article ; Online: Genetic controls and cellular behaviors in branching morphogenesis of the renal collecting system.

Costantini, Frank

Wiley interdisciplinary reviews. Developmental biology

2011 Volume 1, Issue 5, Page(s) 693–713

Abstract: The mammalian kidney, which at maturity contains thousands of nephrons joined to a highly branched collecting duct (CD) system, is an important model system for studying the development of a complex organ. Furthermore, congenital anomalies of the kidney ... ...

Abstract	The mammalian kidney, which at maturity contains thousands of nephrons joined to a highly branched collecting duct (CD) system, is an important model system for studying the development of a complex organ. Furthermore, congenital anomalies of the kidney and urinary tract, often resulting from defects in ureteric bud branching morphogenesis, are relatively common human birth defects. Kidney development is initiated by interactions between the nephric duct and the metanephric mesenchyme, leading to the outgrowth and repeated branching of the ureteric bud epithelium, which gives rise to the entire renal CD system. Meanwhile, signals from the ureteric bud induce the mesenchyme cells to form the nephron epithelia. This review focuses on development of the CD system, with emphasis on the mouse as an experimental system. The major topics covered include the origin and development of the nephric duct, formation of the ureteric bud, branching morphogenesis of the ureteric bud, and elongation of the CDs. The signals, receptors, transcription factors, and other regulatory molecules implicated in these processes are discussed. In addition, our current knowledge of cellular behaviors that are controlled by these genes and underlie development of the collecting system is reviewed.
MeSH term(s)	Animals ; Epithelium/growth & development ; Epithelium/metabolism ; Gene Expression Regulation, Developmental ; Humans ; Kidney/growth & development ; Kidney/metabolism ; Mesoderm/growth & development ; Mesoderm/metabolism ; Mice ; Nephrons/growth & development ; Nephrons/metabolism ; Organogenesis ; Ureter/growth & development ; Ureter/metabolism ; Urinary Tract/growth & development ; Urinary Tract/metabolism
Language	English
Publishing date	2011-06-22
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ISSN	1759-7692
ISSN (online)	1759-7692
DOI	10.1002/wdev.52
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Article ; Online: GDNF/Ret signaling and renal branching morphogenesis: From mesenchymal signals to epithelial cell behaviors.

Costantini, Frank

Organogenesis

2011 Volume 6, Issue 4, Page(s) 252–262

Abstract: Signaling by GDNF through the Ret receptor tyrosine kinase is required for the normal growth and morphogenesis of the ureteric bud (UB) during kidney development. Recent studies have sought to understand the precise role of Ret signaling in this process, ...

Abstract	Signaling by GDNF through the Ret receptor tyrosine kinase is required for the normal growth and morphogenesis of the ureteric bud (UB) during kidney development. Recent studies have sought to understand the precise role of Ret signaling in this process, and the specific responses of UB cells to GDNF. Surprisingly, the requirement for Gdnf and Ret was largely relieved by removing the negative regulator Spry1, revealing unexpected functional overlap between GDNF and FGF10. However, the kidneys that developed without Gdnf/Ret and Spry1 displayed significant branching abnormalities, suggesting a unique role for GDNF in fine-tuning UB branching. GDNF/Ret signaling alters patterns of gene expression in UB tip cells, and one critical event is upregulation of the ETS transcription factors Etv4 and Etv5. Mice lacking Etv4 and Etv5 fail to develop kidneys. Thus, these genes represent key components of a regulatory network downstream of Ret. Studies of chimeric embryos in which a subset of cells lack either Ret, Etv4/5 or Spry1 have revealed an important role for this pathway in cell movement. Ret signaling, via Etv4 and Etv5, promotes competitive cell rearrangements in the nephric duct, in which the cells with the highest level of Ret signaling preferentially migrate to form the first ureteric bud tip.
MeSH term(s)	Animals ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Epithelial Cells/metabolism ; Glial Cell Line-Derived Neurotrophic Factor/metabolism ; Kidney/growth & development ; Kidney/metabolism ; Mesenchymal Stem Cells/metabolism ; Mice ; Morphogenesis ; Proto-Oncogene Proteins c-ets/genetics ; Proto-Oncogene Proteins c-ets/metabolism ; Proto-Oncogene Proteins c-ret/genetics ; Proto-Oncogene Proteins c-ret/metabolism ; Signal Transduction ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Ureter/cytology ; Ureter/growth & development
Chemical Substances	DNA-Binding Proteins ; Etv4 protein, mouse ; Etv5 protein, mouse ; Glial Cell Line-Derived Neurotrophic Factor ; Proto-Oncogene Proteins c-ets ; Transcription Factors ; Proto-Oncogene Proteins c-ret (EC 2.7.10.1)
Language	English
Publishing date	2011-01-15
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2159583-5
ISSN	1555-8592 ; 1555-8592
ISSN (online)	1555-8592
ISSN	1555-8592
DOI	10.4161/org.6.4.12680
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Article: Covid-19-Associated Pulmonary Aspergillosis: The Other Side of the Coin.

Costantini, Claudio / van de Veerdonk, Frank L / Romani, Luigina

Vaccines

2020 Volume 8, Issue 4

Abstract: The immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a critical factor in the clinical presentation of COVID-19, which may range from asymptomatic to a fatal, multi-organ disease. A dysregulated immune response not only ... ...

Abstract	The immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a critical factor in the clinical presentation of COVID-19, which may range from asymptomatic to a fatal, multi-organ disease. A dysregulated immune response not only compromises the ability of the host to resolve the viral infection, but may also predispose the individual to secondary bacterial and fungal infections, a risk to which the current therapeutic immunomodulatory approaches significantly contribute. Among the secondary infections that may occur in COVID-19 patients, coronavirus-associated pulmonary aspergillosis (CAPA) is emerging as a potential cause of morbidity and mortality, although many aspects of the disease still remain unresolved. With this opinion, we present the current view of CAPA and discuss how the same mechanisms that underlie the dysregulated immune response in COVID-19 increase susceptibility to
Language	English
Publishing date	2020-12-01
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines8040713
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Article ; Online: Mosaic analysis of cell rearrangements during ureteric bud branching in dissociated/reaggregated kidney cultures and in vivo.

Leclerc, Kevin / Costantini, Frank

Developmental dynamics : an official publication of the American Association of Anatomists

2016 Volume 245, Issue 4, Page(s) 483–496

Abstract: Background: Cell rearrangements mediated by GDNF/Ret signaling underlie the formation of the ureteric bud (UB) tip domain during kidney development. Whether FGF signaling also influences these rearrangements is unknown. Chimeric embryos are a powerful ... ...

Abstract	Background: Cell rearrangements mediated by GDNF/Ret signaling underlie the formation of the ureteric bud (UB) tip domain during kidney development. Whether FGF signaling also influences these rearrangements is unknown. Chimeric embryos are a powerful tool for examining the genetic controls of cellular behaviors, but generating chimeras by traditional methods is expensive and laborious. Dissociated fetal kidney cells can reorganize to form complex structures including branching UB tubules, providing an easier method to generate renal chimeras. Results: Cell behaviors in normal or chimeric kidney cultures were investigated using time-lapse imaging. In Spry1(-/-) ↔ wild-type chimeras, cells lacking Spry1 (a negative regulator of Ret and FGF receptor signaling) preferentially occupied the UB tips, as previously observed in traditional chimeras, thus validating this experimental system. In Fgfr2(UB-/-) ↔ wild-type chimeras, the wild-type cells preferentially occupied the tips. Independent evidence for a role of Fgfr2 in UB tip formation was obtained using Mosaic mutant Analysis with Spatial and Temporal control of Recombination (MASTR). Conclusions: Dissociation and reaggregation of fetal kidney cells of different genotypes, with suitable fluorescent markers, provides an efficient way to analyze cell behaviors in chimeric cultures. FGF/Fgfr2 signaling promotes UB cell rearrangements that form the tip domain, similarly to GDNF/Ret signaling.
MeSH term(s)	Animals ; Cells, Cultured ; Kidney/cytology ; Kidney/metabolism ; Mice ; Mice, Knockout ; Signal Transduction/physiology
Language	English
Publishing date	2016-04
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1102541-4
ISSN	1097-0177 ; 1058-8388
ISSN (online)	1097-0177
ISSN	1058-8388
DOI	10.1002/dvdy.24387
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Article ; Online: The Role of High-Resolution Manometry Prior to and Following Antireflux Surgery: The Padova Consensus.

Salvador, Renato / Pandolfino, John E / Costantini, Mario / Gyawali, C Prakash / Keller, Jutta / Mittal, Sumeet / Roman, Sabine / Savarino, Edoardo Vincenzo / Tatum, Roger / Tolone, Salvatore / Zerbib, Frank / Capovilla, Giovanni / Jain, Anand / Kathpalia, Priya / Provenzano, Luca / Yadlapati, Rena

Annals of surgery

2024

Abstract: Background: In the last two decades the development of high-resolution manometry (HRM) has changed and revolutionized the diagnostic assessment of patients complain foregut symptoms. The role of HRM before and after antireflux procedure remains unclear, ...

Abstract	Background: In the last two decades the development of high-resolution manometry (HRM) has changed and revolutionized the diagnostic assessment of patients complain foregut symptoms. The role of HRM before and after antireflux procedure remains unclear, especially in surgical practice, where a clear understanding of esophageal physiology and hiatus anatomy is essential for optimal outcome of antireflux surgery (ARS). Surgeons and gastroenterologists (GIs) agree that assessing patients following antireflux procedures can be challenging. Although endoscopy and barium-swallow can reveal anatomic abnormalities, physiologic information on HRM allowing insight into the cause of eventually recurrent symptoms could be key to clinical decision making. Method: A multi-disciplinary international working group (14 surgeons and 15 GIs) collaborated to develop consensus on the role of HRM pre- and post- ARS, and to develop a postoperative classification to interpret HRM findings. The method utilized was detailed literature review to develop statements, and the RAND/University of California, Los Angeles Appropriateness Methodology (RAM) to assess agreement with the statements. Only statements with an approval rate >80% or a final ranking with a median score of 7 were accepted in the consensus. The working groups evaluated the role of HRM prior to ARS and the role of HRM following ARS. Conclusion: This international initiative developed by surgeons and GIs together, summarizes the state of our knowledge of the use of HRM pre- and post-ARS. The Padova Classification was developed to facilitate the interpretation of HRM studies of patients underwent ARS.
Language	English
Publishing date	2024-04-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	340-2
ISSN	1528-1140 ; 0003-4932
ISSN (online)	1528-1140
ISSN	0003-4932
DOI	10.1097/SLA.0000000000006297
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Article ; Online: Low nephron endowment increases susceptibility to renal stress and chronic kidney disease.

Good, Pamela I / Li, Ling / Hurst, Holly A / Serrano Herrera, Ileana / Xu, Katherine / Rao, Meenakshi / Bateman, David A / Al-Awqati, Qais / D'Agati, Vivette D / Costantini, Frank / Lin, Fangming

JCI insight

2023 Volume 8, Issue 3

Abstract: Preterm birth results in low nephron endowment and increased risk of acute kidney injury (AKI) and chronic kidney disease (CKD). To understand the pathogenesis of AKI and CKD in preterm humans, we generated potentially novel mouse models with a 30%-70% ... ...

Abstract	Preterm birth results in low nephron endowment and increased risk of acute kidney injury (AKI) and chronic kidney disease (CKD). To understand the pathogenesis of AKI and CKD in preterm humans, we generated potentially novel mouse models with a 30%-70% reduction in nephron number by inhibiting or deleting Ret tyrosine kinase in the developing ureteric bud. These mice developed glomerular and tubular hypertrophy, followed by the transition to CKD, recapitulating the renal pathological changes seen in humans born preterm. We injected neonatal mice with gentamicin, a ubiquitous nephrotoxic exposure in preterm infants, and detected more severe proximal tubular injury in mice with low nephron number compared with controls with normal nephron number. Mice with low nephron number had reduced proliferative repair with more rapid development of CKD. Furthermore, mice had more profound inflammation with highly elevated levels of MCP-1 and CXCL10, produced in part by damaged proximal tubules. Our study directly links low nephron endowment with postnatal renal hypertrophy, which in this model is maladaptive and results in CKD. Underdeveloped kidneys are more susceptible to gentamicin-induced AKI, suggesting that AKI in the setting of low nephron number is more severe and further increases the risk of CKD in this vulnerable population.
MeSH term(s)	Animals ; Female ; Humans ; Mice ; Acute Kidney Injury/pathology ; Gentamicins ; Hypertrophy/pathology ; Infant, Premature ; Kidney/pathology ; Nephrons/pathology ; Premature Birth/pathology ; Renal Insufficiency, Chronic/pathology
Chemical Substances	Gentamicins
Language	English
Publishing date	2023-02-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.161316
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Article ; Online: Covid-19-Associated Pulmonary Aspergillosis

Claudio Costantini / Frank L. van de Veerdonk / Luigina Romani

Vaccines, Vol 8, Iss 713, p

The Other Side of the Coin

2020 Volume 713

Abstract	The immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a critical factor in the clinical presentation of COVID-19, which may range from asymptomatic to a fatal, multi-organ disease. A dysregulated immune response not only compromises the ability of the host to resolve the viral infection, but may also predispose the individual to secondary bacterial and fungal infections, a risk to which the current therapeutic immunomodulatory approaches significantly contribute. Among the secondary infections that may occur in COVID-19 patients, coronavirus-associated pulmonary aspergillosis (CAPA) is emerging as a potential cause of morbidity and mortality, although many aspects of the disease still remain unresolved. With this opinion, we present the current view of CAPA and discuss how the same mechanisms that underlie the dysregulated immune response in COVID-19 increase susceptibility to Aspergillus infection. Likewise, resorting to endogenous pathways of immunomodulation may not only restore immune homeostasis in COVID-19 patients, but also reduce the risk for aspergillosis. Therefore, CAPA represents the other side of the coin in COVID-19 and our advances in the understanding and treatment of the immune response in COVID-19 should represent the framework for the study of CAPA.
Keywords	COVID-19 ; Aspergillus ; anakinra ; Aryl Hydrocarbon Receptor ; thymosin alpha 1 ; Medicine ; R
Subject code	610
Language	English
Publishing date	2020-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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