High-throughput phenotypic screening of the human spermatozoon

Abstract

Despite recent advances in male reproductive health research, there remain many elements of male (in)fertility where our understanding is incomplete. Consequently, diagnostic tools and treatments for men with sperm dysfunction, other than medically assisted reproduction, are limited. On the other hand, the gaps in our knowledge of the mechanisms which underpin sperm function have hampered the development of male non-hormonal contraceptives. The study of mature spermatozoa is inherently difficult. They are a unique and highly specialised cell type which does not actively transcribe or translate proteins and cannot be cultured for long periods of time or matured in vitro. One, large scale, approach to both increasing understanding of sperm function, and the discovery and development of compounds that can modulate sperm function, is to directly observe responses to compounds with phenotypic screening techniques. These target agnostic approaches can be developed into high-throughput screening platforms with the potential to drastically increase advances in the field. Here we discuss the rationale and development of high-throughput phenotypic screening platforms for mature human spermatozoa, and the multiple potential applications these present, as well as the current limitations and leaps in our understanding and capabilities needed to overcome them. Further development and use of these technologies could lead to the identification of compounds which positively or negatively affect sperm cell motility or function, or novel platforms for toxicology or environmental chemical testing among other applications. Ultimately, each of these potential applications is also likely to increase understanding within the field of sperm biology.

Sperm Cell Painting: A Mechanism Driven Approach for Drug Discovery in Human Spermatozoa

“…To begin to address these gaps in understanding, we have recently developed a high-throughput phenotypic screening platform in human sperm and used it to screen for compounds that either both decrease [1] and enhance [2] sperm motility. This platform presents opportunities for target-agnostic screening which can be adapted for screening of specific sperm functions (reviewed in Johnston et al ., 2021 [3]). Target agnostic phenotypic assays provide a powerful, unbiased approach to drug discovery, but require substantial downstream follow-up of any hits to identify targets and assess the opportunity for further compound development.…”

Male contraceptive development: A medicinal chemistry perspective

“…Therefore, phenotypic screening is a rational and pragmatic approach for contraceptive development. [33] There are several phenotypes of human sperm that may be suitable to investigate. One key phenotype associated with male infertility is reduced motility.…”

An Image-Based High-Throughput and High-Content Drug Screening Method Based on Microarray and Expansion Microscopy

“…3−7 Traditional drug screening is based on multiwell plates, which consume many cells, mediums, and drugs. 4,8,9 In recent years, microarrays began to be extensively developed as a better alternative due to the increased assay throughput and the reduced reagent consumption induced by miniaturiza-tion. 10−15 Zhang et al developed a superhydrophobic microwell array chip (SMARchip).…”

“…Time cost, accessibility, throughput, and content richness are the primary considerations in the evaluation of drug screening methods. Many efforts have been made to optimize the drug screening system. − Traditional drug screening is based on multiwell plates, which consume many cells, mediums, and drugs. ,, In recent years, microarrays began to be extensively developed as a better alternative due to the increased assay throughput and the reduced reagent consumption induced by miniaturization. − Zhang et al . developed a superhydrophobic microwell array chip (SMARchip) .…”