Historical human remains can raise questions at the intersection of palaeopathology, forensic genetics, and identity research. This study discusses the case of a male skeleton with pronounced short stature (1.31 m) and features suggestive of severe skeletal dysplasia, whose body was transferred to the Institute of Pathology in Munich in 1883. The individual was identified as Carl Mühlbacher, a Bavarian man who died while imprisoned, accused of fatally stabbing his mother. Based on bone morphology, rhizomelic dysplasia of the Ain-Naz type was initially considered a plausible diagnosis. However, a targeted evaluation of the reported mutation in the GNPNAT1 gene did not support this hypothesis. Consequently, the remains underwent shotgun sequencing (SGS) to screen for the underlying genetic cause across a broader set of over 500 genes associated with skeletal dysplasia. 

In parallel, we explored the forensic potential of the dataset by integrating documentary evidence with phenotype- and genealogy-oriented DNA analyses. Historical police records describing the individual during imprisonment reported grey eyes and blond-reddish hair. We successfully typed 23 SNPs associated with eye colour, almost 3000 with hair colour and 25 with eyebrow colour, 32 with hair shape and over 2000 with male pattern baldness. Using different prediction models, we consistently arrived at a prediction of brown eyes, brown straight hair, dark eyebrows, and a predisposition to baldness. For identification purposes, we extracted from the SGS data the mitogenome, Y-chromosomal phylogenetic and genealogical SNPs, common autosomal identity SNPs, as well as autosomal and Y-chromosomal STRs. To the extent possible, concordant genotypes and haplotypes were confirmed by comparison with MPS and CE-based data. 

Additionally, a subset of the SGS data (ca. 1.3M SNPs) as well as independently obtained Kintelligence profile were uploaded to GEDmatch PRO™ to investigate whether genetic genealogy could provide identity-relevant leads. Genealogical evaluation identified several present-day individuals with family lineages tracing back to Germany, including families from Carl’s region of origin. The inferred relationships suggested a shared ancestral line through his grandparental or great-grandparental generation, consistent with lineages extending back to the seventeenth century. The further genealogical work is ongoing. 

Together, these findings illustrate the value of SGS as a bridge between palaeopathology, forensics and historical identification. Such combined applications remain uncommon, making this case a compelling example of how forensic genomics can contribute to unresolved historical questions.

Authors

• Marta Diepenbroek (Institute of Legal Medicine, Ludwig Maximilian University of Munich, Germany)
• Marie-Louise Kampmann (Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark)
• Jeppe Dyrberg Andersen (Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark)
• Lotte Sprangers (Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark)
• Brando Poggiali (Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark)
• Arwin Ralf (Department of Pathology and Clinical Bioinformatics, Erasmus MC University Medical Center Rotterdam, Netherlands)
• Manfred Kayser (Department of Pathology and Clinical Bioinformatics, Erasmus MC University Medical Center Rotterdam, Netherlands)
• Katja Anslinger (Institute of Legal Medicine, Ludwig Maximilian University of Munich, Germany)
• Andreas Nerlich (Institute of Legal Medicine, Ludwig Maximilian University of Munich, Germany)