Acquired skewing of lyonization remains stable for a prolonged period in healthy blood donors

ABSTRACT The pattern of X-chromosome inactivation (XCIP), or Lyonization, can be used to distinguish monoclonal from polyclonal cell populations in females. However, a skewed XCIP exists in

hematopoietic cells in approximately 40% of healthy elderly females, interfering with interpretation of clonality assays. In hematopoiesis, an active stem cell pool is assumed to be present

within a larger population of inactive stem cells, with a continuous exchange of cells between the two compartments. The assumption that the active stem cell pool size decreases with age may

explain the phenomenon of acquired skewing occurring by chance and predicts the XCIP of this population to fluctuate. This fluctuation should be reflected in the XCIP of peripheral

granulocytes. We examined the XCIP for fluctuations in time in peripheral granulocytes, monocytes and T cells of young, middle-aged and elderly healthy females. We used an optimized HUMARA

PCR assay that eliminates unbalanced DNA amplification. We found no fluctuations in XCIP in any age group in up to 18 months follow-up. We conclude that acquired skewing arises gradually in

life without fluctuations in XCIP and that analysis at multiple time points cannot distinguish monoclonal hematopoiesis from normal, skewed hematopoiesis. Access through your institution Buy

or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS Access through your institution Subscribe to this journal Receive 12 print issues and

online access $259.00 per year only $21.58 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to local taxes

which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY

OTHERS SKEWNESS OF X-CHROMOSOME INACTIVATION INCREASES WITH AGE AND VARIES ACROSS BIRTH COHORTS IN ELDERLY DANISH WOMEN Article Open access 22 February 2021 DISTINCTION OF LYMPHOID AND

MYELOID CLONAL HEMATOPOIESIS Article 18 October 2021 SHARED AND DISTINCT GENETIC ETIOLOGIES FOR DIFFERENT TYPES OF CLONAL HEMATOPOIESIS Article Open access 08 September 2023 REFERENCES *

Lyon MF . Gene action in the X-chromosome of the mouse (_mus musculus_ L.) _Nature_ 1961 190: 372–373 Article  CAS  PubMed  Google Scholar  * Vogelstein B, Fearon ER, Hamilton SR, Feinberg

AP . Use of restriction fragment length polymorphisms to determine the clonal origin of human tumors _Science_ 1985 227: 642–645 Article  CAS  PubMed  Google Scholar  * Gale RE, Wainscoat JS

. Clonal analysis using X-linked DNA polymorphisms _Br J Haematol_ 1993 85: 2–8 Article  CAS  PubMed  Google Scholar  * Allen RC, Zoghbi HY, Moseley AB, Rosenblatt HM, Belmont JW .

Methylation of _Hpa_II and _Hha_I sites near the polymorphic CAG repeat in the human androgen-receptor gene correlates with X chromosome inactivation _Am J Hum Genet_ 1992 51: 1229–1239 CAS

  PubMed  PubMed Central  Google Scholar  * Mutter GL, Boynton KA . PCR bias in amplification of androgen receptor alleles, a trinucleotide repeat marker used in clonality studies _Nucleic

Acids Res_ 1995 23: 1411–1418 Article  CAS  PubMed  PubMed Central  Google Scholar  * Gale RE, Mein CA, Linch DC . Quantification of X-chromosome inactivation patterns in haematological

samples using the DNA PCR-based HUMARA assay _Leukemia_ 1996 10: 362–367 CAS  PubMed  Google Scholar  * Busque L, Mio R, Mattioli J, Brais E, Blais N, Lalonde Y, Maragh M, Gilliland DG .

Nonrandom X-inactivation patterns in normal females: lyonization ratios vary with age _Blood_ 1996 88: 59–65 CAS  PubMed  Google Scholar  * Champion KM, Gilbert JG, Asimakopoulos FA,

Hinshelwood S, Green AR . Clonal haemopoiesis in normal elderly women: implications for the myeloproliferative disorders and myelodysplastic syndromes _Br J Haematol_ 1997 97: 920–926

Article  CAS  PubMed  Google Scholar  * Gale RE, Fielding AK, Harrison CN, Linch DC . Acquired skewing of X-chromosome inactivation patterns in myeloid cells of the elderly suggests

stochastic clonal loss with age _Br J Haematol_ 1997 98: 512–519 Article  CAS  PubMed  Google Scholar  * Tonon L, Bergamaschi G, Dellavecchia C, Rosti V, Lucotti C, Malabarba L, Novella A,

Vercesi E, Frassoni F, Cazzola M . Unbalanced X-chromosome inactivation in haemopoietic cells from normal women _Br J Haematol_ 1998 102: 996–1003 Article  CAS  PubMed  Google Scholar  *

Nakahara Y, Suzuki H, Ohashi H, Hatano S, Tomita A, Kinoshita T, Murate T, Saito H, Hotta T . Clonality analysis of granulocytes and T lymphocytes in healthy females by the PCR-based HUMARA

method _Int J Hematol_ 1999 69: 237–243 CAS  PubMed  Google Scholar  * Wiggans RG, Jacobson RJ, Fialkow PJ, Woolley PV, Macdonald JS, Schein PS . Probable clonal origin of acute myeloblastic

leukemia following radiation and chemotherapy of colon cancer _Blood_ 1978 52: 659–663 CAS  PubMed  Google Scholar  * Gale RE, Bunch C, Moir DJ, Patterson KG, Goldstone AH, Linch DC .

Demonstration of developing myelodysplasia/acute myeloid leukaemia in haematologically normal patients after high-dose chemotherapy and autologous bone marrow transplantation using

X-chromosome inactivation patterns _Br J Haematol_ 1996 93: 53–58 Article  CAS  PubMed  Google Scholar  * Fialkow PJ, Faguet GB, Jacobson RJ, Vaidya K, Murphy S . Evidence that essential

thrombocythemia is a clonal disorder with origin in a multipotent stem cell _Blood_ 1981 58: 916–919 CAS  PubMed  Google Scholar  * Anger B, Janssen JW, Schrezenmeier H, Hehlmann R, Heimpel

H, Bartram CR . Clonal analysis of chronic myeloproliferative disorders using X-linked DNA polymorphisms _Leukemia_ 1990 4: 258–261 CAS  PubMed  Google Scholar  * Cheshier SH, Morrison SJ,

Liao X, Weissman IL . _In vivo_ proliferation and cell cycle kinetics of long-term self-renewing hematopoietic stem cells _Proc Natl Acad Sci USA_ 1999 96: 3120–3125 Article  CAS  PubMed 

PubMed Central  Google Scholar  * Abkowitz JL, Golinelli D, Harrison DE, Guttorp P . _In vivo_ kinetics of murine hemopoietic stem cells _Blood_ 2000 96: 3399–3405 CAS  PubMed  Google

Scholar  * Wang JC, Doedens M, Dick JE . Primitive human hematopoietic cells are enriched in cord blood compared with adult bone marrow or mobilized peripheral blood as measured by the

quantitative _in vivo_ SCID-repopulating cell assay _Blood_ 1997 89: 3919–3924 CAS  PubMed  Google Scholar  * Abkowitz JL, Taboada M, Shelton GH, Catlin SN, Guttorp P, Kiklevich JV . An X

chromosome gene regulates hematopoietic stem cell kinetics _Proc Natl Acad Sci USA_ 1998 95: 3862–3866 Article  CAS  PubMed  PubMed Central  Google Scholar  * Christensen K, Kristiansen M,

Hagen-Larsen H, Skytthe A, Bathum L, Jeune B, Andersen-Ranberg K, Vaupel JW, Orstavik KH . X-linked genetic factors regulate hematopoietic stem-cell kinetics in females _Blood_ 2000 95:

2449–2451 CAS  PubMed  Google Scholar  * Vickers MA, McLeod E, Spector TD, Wilson IJ . Assessment of mechanism of acquired skewed X inactivation by analysis of twins _Blood_ 2001 97:

1274–1281 Article  CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by a grant from the Dutch Cancer Society (NKB). AUTHOR INFORMATION AUTHORS AND

AFFILIATIONS * Central Hematology Laboratory, University Medical Center Nijmegen, Nijmegen, The Netherlands JP van Dijk, L Heuver, E Stevens-Linders, JH Jansen & EJBM Mensink *

Department of Hematology, University Medical Center Nijmegen, Nijmegen, The Netherlands RAP Raymakers & T de Witte Authors * JP van Dijk View author publications You can also search for

this author inPubMed Google Scholar * L Heuver View author publications You can also search for this author inPubMed Google Scholar * E Stevens-Linders View author publications You can also

search for this author inPubMed Google Scholar * JH Jansen View author publications You can also search for this author inPubMed Google Scholar * EJBM Mensink View author publications You

can also search for this author inPubMed Google Scholar * RAP Raymakers View author publications You can also search for this author inPubMed Google Scholar * T de Witte View author

publications You can also search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE van Dijk, J., Heuver, L.,

Stevens-Linders, E. _et al._ Acquired skewing of Lyonization remains stable for a prolonged period in healthy blood donors. _Leukemia_ 16, 362–367 (2002).

https://doi.org/10.1038/sj.leu.2402379 Download citation * Received: 15 June 2001 * Accepted: 14 November 2001 * Published: 04 March 2002 * Issue Date: 01 March 2002 * DOI:

https://doi.org/10.1038/sj.leu.2402379 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not

currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * hematopoiesis * Lyonization * skewing * HUMARA