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4103 Distribution of Chromosome-Arm Specific Telomere Length in Patients with Telomere Biology Disorders

Program: Oral and Poster Abstracts
Session: 509. Bone Marrow Failure and Cancer Predisposition Syndromes: Congenital: Poster III
Hematology Disease Topics & Pathways:
Research, Fundamental Science, Acute Myeloid Malignancies, AML, artificial intelligence (AI), Translational Research, epidemiology, Bone Marrow Failure Syndromes, Inherited Marrow Failure Syndromes, CHIP, assays, Clinical Research, health outcomes research, Genetic Disorders, genomics, bioinformatics, hematopoiesis, Diseases, real-world evidence, registries, computational biology, Myeloid Malignancies, Biological Processes, emerging technologies, molecular biology, Technology and Procedures, gene editing, Minimal Residual Disease , machine learning, molecular testing, natural language processing, omics technologies
Monday, December 11, 2023, 6:00 PM-8:00 PM

Alejandro Ferrer, PhD1, Zachary D Stephens, Ph.D.2*, Susan M. Geyer, PhD3, Rachel A Simon4*, Terra L. Lasho, PhD5, Christy M Finke5*, Brittany Burnap1*, Lindsay A Mulvihill, CCRP6*, Tammy M McAllister, M.A.6*, Carolyn R. Rohrer Vitek, Ed.D. / M.S.6*, Konstantinos N Lazaridis1*, Angita Jain7*, Eric W Klee, PhD8*, Jean-Pierre A Kocher, Ph.D.8* and Mrinal M. Patnaik, MD, MBBS4

1Mayo Clinic, Rochester, MN
2Department of Computational Biology, Mayo Clinic, Rochester, MN
3Alliance Statistics and Data Management Center, Mayo Clinic, Rochester, MN
4Division of Hematology, Mayo Clinic, Rochester, MN
5Division of Hematology, Department of Medicine, Mayo Clinic, Rochester, MN
6Center for Individualized Medicine, Mayo Clinic, Rochester, MN
7Mayo Clinic, Jacksonville, FL
8Computational Biology, Mayo Clinic, Rochester, MN

Introduction: Telomere biology disorders (TBD) are multisystem diseases often associated with bone marrow failure (BMF) and pulmonary fibrosis. TBD are clinically diagnosed when mean telomere lengths (TL) in blood cells measured by clinical flowFISH assays fall <1st percentile, in comparison to age-matched reference controls. This information has limitations, e.g. TL in blood cells may not represent TL in other organs (e.g. lung, liver) and moreover, individuals with high clinical suspicion of TBD can show borderline average TL which complicates their diagnosis. The likely reason for this is that cell senescence can be triggered by shortening of a subset, or even a single chromosome arm telomere, a situation that may be missed when measuring average TL. Previous approaches to measure individual chromosome-arm telomere length (CA-TL) are only available for a small number of chromosome arms or can only estimate the number of shortened telomeres without matching them to specific chromosome arms. Taking advantage of recent developments in long-read next generation sequencing (NGS) and the availability of the recently published telomere-to-telomere (T2T) reference human genome, our group developed Telogator, (Stephens Z. et al, 2022) a bioinformatic tool that estimates CA-TL for most chromosome arms using long-read NGS data. Here we use Telogator to describe for the first time the CA-TL pattern in TBD patients in comparison with a control non-TBD cohort.

Methods: TBD patients were identified through the Pre-Myeloid and Bone Marrow Failure Specialty Clinic at Mayo Clinic. All patients underwent flowFISH and genetic testing. Age-matched non-TBD patients identified at the Center for Individualized Medicine (Mayo Clinic) were used as reference controls. Informed consent was obtained from all individuals with approval from the Mayo Clinic Institutional Review Board. DNA from peripheral blood mononuclear cells was sequenced using PacBio Sequel II following the manufacturer indications for HiFi sequencing. BAM files were aligned to the T2T reference and CA-TL was calculated using Telogator as described in Stephens Z. et al. 2022. Average TL, CA-TL, and percent of short telomeres (both defined as <5 kb and <3.2 kb) were calculated. Comparisons of average TL and percent of short telomeres were performed using t-test, while Wilcoxon rank-sum test was used to compare CA-TL.

Results: Four TBD patients [median age = 58.5 (52-60), 50% female] and 6 non-TBD controls [Median age = 56 (50-63), 33% female] were included in the analysis. All TBD patients presented average TL below the 1st centile in both lymphocytes and granulocytes, and TBD-related phenotype (100% with pulmonary fibrosis, 50% with BMF, 25% with liver fibrosis). Three had a pathogenic variant in a TBD related genes [2 in TERT (NM_198253.3: c.2030G>A, p.(Gly677Asp) and c.2515_2533del, p.(Thr839Alafs*29)) and 1 in NHP2 (NM_017838.3: c.459_460delAT; p.(X154Argext29))].

The average TL calculated by Telogator showed high correlation with flowFISH values (r2 = 0.9883, p = 0.0059) and was lower in TBD patients than controls (p = 0.0012). Next, when comparing individual CA-TL between groups, only half of CA-TL was statistically different in the TBD group compared to controls (1p, 3q, 4p, 5p, 5q, 6q, 7p, 7q, 8q, 9p, 9q, 10q, 11p, 12q, 12q, 16p, 16q, 18q, 19p, 19q, 20p, 20q, 21q – Figure 1). The percentage of telomeres <5kb and <3 kb was significantly increased in TBD patients compared to controls (p= 0.014 and 0.0333, respectively).

Discussion: Using a novel informatic tool, we replicate previous results obtained using classic approaches and were able to expand on these studies by measuring specific CA-TL in TBD patients. We identified statistically significant differences at the CA-TL level in these patients when compared to age-matched controls, while other CA-TL were not statistically different. Future studies and the inclusion of additional TBD patients will help define the clinical relevance of the differential CA-TL shortening and its prevalence among TBD patients. While limited by the small number of samples in the study, we show the feasibility of using this approach to study TL with unprecedented resolution and show that Telogator offers important advantages compared to classic methods to study telomere biology that can be used in other relevant fields like aging and/or cancer. Our results pave the way to advance the mechanistic understanding of TBD.

Disclosures: Klee: Undiganozied Disease Network Int'l: Membership on an entity's Board of Directors or advisory committees; Helix Population Genomics: Patents & Royalties: Helix Population Genomics. Patnaik: Epigenetix: Research Funding; Kura Oncology: Research Funding; CTI Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; StemLine: Research Funding.

*signifies non-member of ASH