FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans

Qiumei Du; Larry K. Huynh; Fatma Coskun; Erika Molina; Matthew A. King; Prithvi Raj; Shaheen Khan; Igor Dozmorov; Christine M. Seroogy; Christian A. Wysocki; Grace T. Padron; Tyler R. Yates; M. Louise Markert; M. Teresa de la Morena; Nicolai S.C. van Oers

doi:10.1172/JCI127565

FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans

Qiumei Du, Larry K. Huynh, Fatma Coskun, Erika Molina, Matthew A. King, Prithvi Raj, Shaheen Khan, Igor Dozmorov, Christine M. Seroogy, Christian A. Wysocki, Grace T. Padron, Tyler R. Yates, M. Louise Markert, M. Teresa de la Morena, Nicolai S.C. van Oers

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

We report on 2 patients with compound heterozygous mutations in forkhead box N1 (FOXN1), a transcription factor essential for thymic epithelial cell (TEC) differentiation. TECs are critical for T cell development. Both patients had a presentation consistent with T^–/loB+NK⁺ SCID, with normal hair and nails, distinct from the classic nude/SCID phenotype in individuals with autosomal-recessive FOXN1 mutations. To understand the basis of this phenotype and the effects of the mutations on FOXN1, we generated mice using CRISPR-Cas9 technology to genocopy mutations in 1 of the patients. The mice with the Foxn1 compound heterozygous mutations had thymic hypoplasia, causing a T–B+NK+ SCID phenotype, whereas the hair and nails of these mice were normal. Characterization of the functional changes due to the Foxn1 mutations revealed a 5–amino acid segment at the end of the DNA-binding domain essential for the development of TECs but not keratinocytes. The transcriptional activity of this Foxn1 mutant was partly retained, indicating a region that specifies TEC functions. Analysis of an additional 9 FOXN1 mutations identified in multiple unrelated patients revealed distinct functional consequences contingent on the impact of the mutation on the DNA-binding and transactivation domains of FOXN1.

Original language	English (US)
Pages (from-to)	4724-4738
Number of pages	15
Journal	Journal of Clinical Investigation
Volume	129
Issue number	11
DOIs	https://doi.org/10.1172/JCI127565
State	Published - Nov 1 2019
Externally published	Yes

ASJC Scopus subject areas

Medicine(all)

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10.1172/JCI127565

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Du, Q., Huynh, L. K., Coskun, F., Molina, E., King, M. A., Raj, P., Khan, S., Dozmorov, I., Seroogy, C. M., Wysocki, C. A., Padron, G. T., Yates, T. R., Louise Markert, M., Teresa de la Morena, M., & van Oers, N. S. C. (2019). FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans. Journal of Clinical Investigation, 129(11), 4724-4738. https://doi.org/10.1172/JCI127565

Du, Q, Huynh, LK, Coskun, F, Molina, E, King, MA, Raj, P, Khan, S, Dozmorov, I, Seroogy, CM, Wysocki, CA, Padron, GT, Yates, TR, Louise Markert, M, Teresa de la Morena, M & van Oers, NSC 2019, 'FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans', Journal of Clinical Investigation, vol. 129, no. 11, pp. 4724-4738. https://doi.org/10.1172/JCI127565

@article{7843e1f1707549c68131cc30e2f04c9f,

title = "FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans",

abstract = "We report on 2 patients with compound heterozygous mutations in forkhead box N1 (FOXN1), a transcription factor essential for thymic epithelial cell (TEC) differentiation. TECs are critical for T cell development. Both patients had a presentation consistent with T–/loB+NK+ SCID, with normal hair and nails, distinct from the classic nude/SCID phenotype in individuals with autosomal-recessive FOXN1 mutations. To understand the basis of this phenotype and the effects of the mutations on FOXN1, we generated mice using CRISPR-Cas9 technology to genocopy mutations in 1 of the patients. The mice with the Foxn1 compound heterozygous mutations had thymic hypoplasia, causing a T–B+NK+ SCID phenotype, whereas the hair and nails of these mice were normal. Characterization of the functional changes due to the Foxn1 mutations revealed a 5–amino acid segment at the end of the DNA-binding domain essential for the development of TECs but not keratinocytes. The transcriptional activity of this Foxn1 mutant was partly retained, indicating a region that specifies TEC functions. Analysis of an additional 9 FOXN1 mutations identified in multiple unrelated patients revealed distinct functional consequences contingent on the impact of the mutation on the DNA-binding and transactivation domains of FOXN1.",

author = "Qiumei Du and Huynh, {Larry K.} and Fatma Coskun and Erika Molina and King, {Matthew A.} and Prithvi Raj and Shaheen Khan and Igor Dozmorov and Seroogy, {Christine M.} and Wysocki, {Christian A.} and Padron, {Grace T.} and Yates, {Tyler R.} and {Louise Markert}, M. and {Teresa de la Morena}, M. and {van Oers}, {Nicolai S.C.}",

note = "Funding Information: We would like to thank Angela Mobley and Kayla Lopez from the flow cytometry core at UT Southwestern Medical Center for assistance with TEC sorting and flow analyses. We appreciate the suggestions and help provided by members of the van Oers laboratory. We are thankful for the statistical analysis that was suggested and/or provided by Lindsay Cowell (Department of Bio-informatics, UT Southwestern Medical Center). Caitlyn Braisch from Ondine Cleaver{\textquoteright}s laboratory at UT Southwestern Medical Center provided image acquisition assistance. We are grateful for the advice provided by Dong-Ming Su (University of North Texas Health Sciences Center) and Nancy Manley (Department of Genetics, University of Georgia). James Richardson, John Shelton, and staff from the Cardiology Core of UT Southwestern Medical Center provided help with thymic tissue and skin processing. The various Foxn1-targeted mice were generated with the help of Robert Hammer of the Transgenic and Knockout Core at UT Southwestern Medical Center. Salius Z{\"u}klys and Georg Hollander (University Children{\textquoteright}s Hospital and University of Basel, Basel, Switzerland) provided the β5t transcriptional reporter construct. We further acknowledge the following physicians who provided genetic and clinical data on deidentified individuals with diverse FOXN1 mutations. Patient information was provided to M. Louise Markert at Duke University (Durham, North Carolina, USA). The physicians included Jennifer Heimall (Children{\textquoteright}s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA); Elena Perez (Allergy Associates of the Palm Beaches, North Palm Beach Florida, USA); John Bohnsack and Karin Chen (University of Utah Health, Salt Lake City, Utah, USA); Challice Bonifant (C.S. Mott Children{\textquoteright}s Hospital, Ann Arbor, Michigan, USA); Hana Niebur (Children{\textquoteright}s Hospital and Medical Center, Omaha, Nebraska, USA); Jordan Abbott and Erwin W. Gelfand (National Jewish Health, Denver, Colorado, USA); Melissa E. Elder (University of Florida, Gaines-ville, Florida, USA); Lisa Forbes and Jacob J.H. Bleesing (University of Cincinnati, Cincinnati, Ohio, USA); David Buchbinder (Children{\textquoteright}s Hospital of Orange County, Orange, California, USA); Melanie Hankewzcz (Hackensack Meridian, Hackensack, New Jersey, USA); Chaim Roifman (Sick Kids, Toronto, Canada); Evan B. Shereck (Oregon Health and Science University, Portland, Oregon, USA); Megan Cooper (St. Louis Children{\textquoteright}s Hospital, St. Louis, Missouri, USA); Lisa Bartnikas (Boston Children{\textquoteright}s Hospital, Boston, Massachusetts, USA); Rebecca H. Buckley (Duke University Medical Center, Durham,North Carolina, USA); Jennifer Leiding (University of South Florida, Saint Petersburg, Florida, USA); Anthony Hayward (Brown University, Providence, Rhode Island, USA); Benjamin T. Prince (Nationwide Children{\textquoteright}s Hospital, Columbus, Ohio, USA); Neena Kapoor (Children{\textquoteright}s Hospital of Los Angeles, Los Angeles, California, USA); Mark T. Vander Lugt (C.S. Mott Children{\textquoteright}s Hospital, Ann Arbor, Michigan, USA); and Tory Quigg (Methodist Children{\textquoteright}s Hospital, San Antonio, Texas, USA). Our work was supported in part by grants from the NIH (R01 AI114523 and R21 AI144140, to NSCVO); Beecherl funds from the Department of Immunology at UT Southwestern Medical Center (to NSCVO); and the Jeffrey Modell Foundation (to MDLM and CAW). Our work was also supported, in part, by grants from the NIH R01 (R01 AI114523, R21 AI144140 NSCVO), and the Jeffrey Modell Foundation (MDLM). Funding Information: We would like to thank Angela Mobley and Kayla Lopez from the flow cytometry core at UT Southwestern Medical Center for assistance with TEC sorting and flow analyses. We appreciate the suggestions and help provided by members of the van Oers laboratory. We are thankful for the statistical analysis that was suggested and/or provided by Lindsay Cowell (Department of Bio-informatics, UT Southwestern Medical Center). Caitlyn Braisch from Ondine Cleaver{\textquoteright}s laboratory at UT Southwestern Medical Center provided image acquisition assistance. We are grateful for the advice provided by Dong-Ming Su (University of North Texas Health Sciences Center) and Nancy Manley (Department of Genetics, University of Georgia). James Richardson, John Shelton, and staff from the Cardiology Core of UT Southwestern Medical Center provided help with thymic tissue and skin processing. The various Foxn1-targeted mice were generated with the help of Robert Hammer of the Transgenic and Knockout Core at UT Southwestern Medical Center. Salius Z{\"u}klys and Georg Hollander (University Children{\textquoteright}s Hospital and University of Basel, Basel, Switzerland) provided the β5t transcriptional reporter construct. We further acknowledge the following physicians who provided genetic and clinical data on deidentified individuals with diverse FOXN1 mutations. Patient information was provided to M. Louise Markert at Duke University (Durham, North Carolina, USA). The physicians included Jennifer Heimall (Children{\textquoteright}s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA); Elena Perez (Allergy Associates of the Palm Beaches, North Palm Beach Florida, USA); John Bohnsack and Karin Chen (University of Utah Health, Salt Lake City, Utah, USA); Challice Bonifant (C.S. Mott Children{\textquoteright}s Hospital, Ann Arbor, Michigan, USA); Hana Niebur (Children{\textquoteright}s Hospital and Medical Center, Omaha, Nebraska, USA); Jordan Abbott and Erwin W. Gelfand (National Jewish Health, Denver, Colorado, USA); Melissa E. Elder (University of Florida, Gainesville, Florida, USA); Lisa Forbes and Jacob J.H. Bleesing (University of Cincinnati, Cincinnati, Ohio, USA); David Buchbinder (Children{\textquoteright}s Hospital of Orange County, Orange, California, USA); Melanie Hankewzcz (Hackensack Meridian, Hackensack, New Jersey, USA); Chaim Roifman (Sick Kids, Toronto, Canada); Evan B. Shereck (Oregon Health and Science University, Portland, Oregon, USA); Megan Cooper (St. Louis Children{\textquoteright}s Hospital, St. Louis, Missouri, USA); Lisa Bartnikas (Boston Children{\textquoteright}s Hospital, Boston, Massachusetts, USA); Rebecca H. Buckley (Duke University Medical Center, Durham,North Carolina, USA); Jennifer Leiding (University of South Florida, Saint Petersburg, Florida, USA); Anthony Hayward (Brown University, Providence, Rhode Island, USA); Benjamin T. Prince (Nationwide Children{\textquoteright}s Hospi- tal, Columbus, Ohio, USA); Neena Kapoor (Children{\textquoteright}s Hospital of Los Angeles, Los Angeles, California, USA); Mark T. Vander Lugt (C.S. Mott Children{\textquoteright}s Hospital, Ann Arbor, Michigan, USA); and Tory Quigg (Methodist Children{\textquoteright}s Hospital, San Antonio, Texas, USA). Our work was supported in part by grants from the NIH (R01 AI114523 and R21 AI144140, to NSCVO); Beecherl funds from the Department of Immunology at UT Southwestern Medical Center (to NSCVO); and the Jeffrey Modell Foundation (to MDLM and CAW). Our work was also supported, in part, by grants from the NIH R01 (R01 AI114523, R21 AI144140 NSCVO), and the Jeffrey Modell Foundation (MDLM). Publisher Copyright: {\textcopyright} 2019, American Society for Clinical Investigation.",

year = "2019",

month = nov,

day = "1",

doi = "10.1172/JCI127565",

language = "English (US)",

volume = "129",

pages = "4724--4738",

journal = "Journal of Clinical Investigation",

issn = "0021-9738",

publisher = "The American Society for Clinical Investigation",

number = "11",

}

TY - JOUR

T1 - FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans

AU - Du, Qiumei

AU - Huynh, Larry K.

AU - Coskun, Fatma

AU - Molina, Erika

AU - King, Matthew A.

AU - Raj, Prithvi

AU - Khan, Shaheen

AU - Dozmorov, Igor

AU - Seroogy, Christine M.

AU - Wysocki, Christian A.

AU - Padron, Grace T.

AU - Yates, Tyler R.

AU - Louise Markert, M.

AU - Teresa de la Morena, M.

AU - van Oers, Nicolai S.C.

N1 - Funding Information: We would like to thank Angela Mobley and Kayla Lopez from the flow cytometry core at UT Southwestern Medical Center for assistance with TEC sorting and flow analyses. We appreciate the suggestions and help provided by members of the van Oers laboratory. We are thankful for the statistical analysis that was suggested and/or provided by Lindsay Cowell (Department of Bio-informatics, UT Southwestern Medical Center). Caitlyn Braisch from Ondine Cleaver’s laboratory at UT Southwestern Medical Center provided image acquisition assistance. We are grateful for the advice provided by Dong-Ming Su (University of North Texas Health Sciences Center) and Nancy Manley (Department of Genetics, University of Georgia). James Richardson, John Shelton, and staff from the Cardiology Core of UT Southwestern Medical Center provided help with thymic tissue and skin processing. The various Foxn1-targeted mice were generated with the help of Robert Hammer of the Transgenic and Knockout Core at UT Southwestern Medical Center. Salius Züklys and Georg Hollander (University Children’s Hospital and University of Basel, Basel, Switzerland) provided the β5t transcriptional reporter construct. We further acknowledge the following physicians who provided genetic and clinical data on deidentified individuals with diverse FOXN1 mutations. Patient information was provided to M. Louise Markert at Duke University (Durham, North Carolina, USA). The physicians included Jennifer Heimall (Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA); Elena Perez (Allergy Associates of the Palm Beaches, North Palm Beach Florida, USA); John Bohnsack and Karin Chen (University of Utah Health, Salt Lake City, Utah, USA); Challice Bonifant (C.S. Mott Children’s Hospital, Ann Arbor, Michigan, USA); Hana Niebur (Children’s Hospital and Medical Center, Omaha, Nebraska, USA); Jordan Abbott and Erwin W. Gelfand (National Jewish Health, Denver, Colorado, USA); Melissa E. Elder (University of Florida, Gaines-ville, Florida, USA); Lisa Forbes and Jacob J.H. Bleesing (University of Cincinnati, Cincinnati, Ohio, USA); David Buchbinder (Children’s Hospital of Orange County, Orange, California, USA); Melanie Hankewzcz (Hackensack Meridian, Hackensack, New Jersey, USA); Chaim Roifman (Sick Kids, Toronto, Canada); Evan B. Shereck (Oregon Health and Science University, Portland, Oregon, USA); Megan Cooper (St. Louis Children’s Hospital, St. Louis, Missouri, USA); Lisa Bartnikas (Boston Children’s Hospital, Boston, Massachusetts, USA); Rebecca H. Buckley (Duke University Medical Center, Durham,North Carolina, USA); Jennifer Leiding (University of South Florida, Saint Petersburg, Florida, USA); Anthony Hayward (Brown University, Providence, Rhode Island, USA); Benjamin T. Prince (Nationwide Children’s Hospital, Columbus, Ohio, USA); Neena Kapoor (Children’s Hospital of Los Angeles, Los Angeles, California, USA); Mark T. Vander Lugt (C.S. Mott Children’s Hospital, Ann Arbor, Michigan, USA); and Tory Quigg (Methodist Children’s Hospital, San Antonio, Texas, USA). Our work was supported in part by grants from the NIH (R01 AI114523 and R21 AI144140, to NSCVO); Beecherl funds from the Department of Immunology at UT Southwestern Medical Center (to NSCVO); and the Jeffrey Modell Foundation (to MDLM and CAW). Our work was also supported, in part, by grants from the NIH R01 (R01 AI114523, R21 AI144140 NSCVO), and the Jeffrey Modell Foundation (MDLM). Funding Information: We would like to thank Angela Mobley and Kayla Lopez from the flow cytometry core at UT Southwestern Medical Center for assistance with TEC sorting and flow analyses. We appreciate the suggestions and help provided by members of the van Oers laboratory. We are thankful for the statistical analysis that was suggested and/or provided by Lindsay Cowell (Department of Bio-informatics, UT Southwestern Medical Center). Caitlyn Braisch from Ondine Cleaver’s laboratory at UT Southwestern Medical Center provided image acquisition assistance. We are grateful for the advice provided by Dong-Ming Su (University of North Texas Health Sciences Center) and Nancy Manley (Department of Genetics, University of Georgia). James Richardson, John Shelton, and staff from the Cardiology Core of UT Southwestern Medical Center provided help with thymic tissue and skin processing. The various Foxn1-targeted mice were generated with the help of Robert Hammer of the Transgenic and Knockout Core at UT Southwestern Medical Center. Salius Züklys and Georg Hollander (University Children’s Hospital and University of Basel, Basel, Switzerland) provided the β5t transcriptional reporter construct. We further acknowledge the following physicians who provided genetic and clinical data on deidentified individuals with diverse FOXN1 mutations. Patient information was provided to M. Louise Markert at Duke University (Durham, North Carolina, USA). The physicians included Jennifer Heimall (Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA); Elena Perez (Allergy Associates of the Palm Beaches, North Palm Beach Florida, USA); John Bohnsack and Karin Chen (University of Utah Health, Salt Lake City, Utah, USA); Challice Bonifant (C.S. Mott Children’s Hospital, Ann Arbor, Michigan, USA); Hana Niebur (Children’s Hospital and Medical Center, Omaha, Nebraska, USA); Jordan Abbott and Erwin W. Gelfand (National Jewish Health, Denver, Colorado, USA); Melissa E. Elder (University of Florida, Gainesville, Florida, USA); Lisa Forbes and Jacob J.H. Bleesing (University of Cincinnati, Cincinnati, Ohio, USA); David Buchbinder (Children’s Hospital of Orange County, Orange, California, USA); Melanie Hankewzcz (Hackensack Meridian, Hackensack, New Jersey, USA); Chaim Roifman (Sick Kids, Toronto, Canada); Evan B. Shereck (Oregon Health and Science University, Portland, Oregon, USA); Megan Cooper (St. Louis Children’s Hospital, St. Louis, Missouri, USA); Lisa Bartnikas (Boston Children’s Hospital, Boston, Massachusetts, USA); Rebecca H. Buckley (Duke University Medical Center, Durham,North Carolina, USA); Jennifer Leiding (University of South Florida, Saint Petersburg, Florida, USA); Anthony Hayward (Brown University, Providence, Rhode Island, USA); Benjamin T. Prince (Nationwide Children’s Hospi- tal, Columbus, Ohio, USA); Neena Kapoor (Children’s Hospital of Los Angeles, Los Angeles, California, USA); Mark T. Vander Lugt (C.S. Mott Children’s Hospital, Ann Arbor, Michigan, USA); and Tory Quigg (Methodist Children’s Hospital, San Antonio, Texas, USA). Our work was supported in part by grants from the NIH (R01 AI114523 and R21 AI144140, to NSCVO); Beecherl funds from the Department of Immunology at UT Southwestern Medical Center (to NSCVO); and the Jeffrey Modell Foundation (to MDLM and CAW). Our work was also supported, in part, by grants from the NIH R01 (R01 AI114523, R21 AI144140 NSCVO), and the Jeffrey Modell Foundation (MDLM). Publisher Copyright: © 2019, American Society for Clinical Investigation.

PY - 2019/11/1

Y1 - 2019/11/1

N2 - We report on 2 patients with compound heterozygous mutations in forkhead box N1 (FOXN1), a transcription factor essential for thymic epithelial cell (TEC) differentiation. TECs are critical for T cell development. Both patients had a presentation consistent with T–/loB+NK+ SCID, with normal hair and nails, distinct from the classic nude/SCID phenotype in individuals with autosomal-recessive FOXN1 mutations. To understand the basis of this phenotype and the effects of the mutations on FOXN1, we generated mice using CRISPR-Cas9 technology to genocopy mutations in 1 of the patients. The mice with the Foxn1 compound heterozygous mutations had thymic hypoplasia, causing a T–B+NK+ SCID phenotype, whereas the hair and nails of these mice were normal. Characterization of the functional changes due to the Foxn1 mutations revealed a 5–amino acid segment at the end of the DNA-binding domain essential for the development of TECs but not keratinocytes. The transcriptional activity of this Foxn1 mutant was partly retained, indicating a region that specifies TEC functions. Analysis of an additional 9 FOXN1 mutations identified in multiple unrelated patients revealed distinct functional consequences contingent on the impact of the mutation on the DNA-binding and transactivation domains of FOXN1.

AB - We report on 2 patients with compound heterozygous mutations in forkhead box N1 (FOXN1), a transcription factor essential for thymic epithelial cell (TEC) differentiation. TECs are critical for T cell development. Both patients had a presentation consistent with T–/loB+NK+ SCID, with normal hair and nails, distinct from the classic nude/SCID phenotype in individuals with autosomal-recessive FOXN1 mutations. To understand the basis of this phenotype and the effects of the mutations on FOXN1, we generated mice using CRISPR-Cas9 technology to genocopy mutations in 1 of the patients. The mice with the Foxn1 compound heterozygous mutations had thymic hypoplasia, causing a T–B+NK+ SCID phenotype, whereas the hair and nails of these mice were normal. Characterization of the functional changes due to the Foxn1 mutations revealed a 5–amino acid segment at the end of the DNA-binding domain essential for the development of TECs but not keratinocytes. The transcriptional activity of this Foxn1 mutant was partly retained, indicating a region that specifies TEC functions. Analysis of an additional 9 FOXN1 mutations identified in multiple unrelated patients revealed distinct functional consequences contingent on the impact of the mutation on the DNA-binding and transactivation domains of FOXN1.

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U2 - 10.1172/JCI127565

DO - 10.1172/JCI127565

M3 - Article

C2 - 31566583

AN - SCOPUS:85074379045

SN - 0021-9738

VL - 129

SP - 4724

EP - 4738

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

IS - 11

ER -

FOXN1 compound heterozygous mutations cause selective thymic hypoplasia in humans

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